The world’s premier 9th Annual Meeting research event

Final Program June 15 – 18, 2011 Metro Convention Centre Toronto, Ontario www.isscr.org

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Letter from ISSCR Dear Colleagues:

On behalf of the International Society for Stem Cell Research, welcome to the ISSCR 9th Annual Meeting at the Metro Toronto Convention Centre. The ISSCR Annual Meeting is the preeminent international forum for stem cell science.

From Wednesday afternoon through Saturday evening, this meeting is packed with the best and brightest the stem cell field has to offer. One hundred eleven speakers from academia and industry around the world will present their latest and most exciting research. Additionally, all posters will be on display for the duration of the meeting, giving you more time than ever to explore the great science in the poster hall. As usual, there are also a number of social and educational activities for junior investigators, students and postdocs, and there are many opportunities for networking at all levels.

This year, the morning prior to the start of the meeting on Wednesday will also be special. In addition to industry- oriented programs and focus sessions highlighting important topics in bioethics and basic science, there will be a satellite symposium honoring the late Ernest McCulloch, one of the true pioneers of stem cell . Admission to the satellite sessions is free to all meeting attendees and we encourage you to participate.

ISSCR is very pleased to be able to honor the memory of one of the giants of our field by introducing the Ernest McCulloch Memorial Lecture here in Toronto. ISSCR will also be introducing two new awards at this meeting, the ISSCR Public Service Award and the McEwen Centre Award for Innovation. You can read about the recipients of these two awards and the ISSCR Outstanding Young Investigator Award in this program.

Judging from advanced registration and abstract submissions, this meeting is on track to be one of the largest and most successful ever for ISSCR. We think you are going to find it as exciting and stimulating as we expect it to be, and are happy to have you here.

Haifan Lin Elaine Fuchs ISSCR Program Chair ISSCR President

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Table of Contents

ISSCR 9th Annual Meeting Co-Sponsor 3 ISSCR Media Partners 36

ISSCR 9th Annual Meeting Supporters 4 – 5 Exhibits and Exhibitor Floor Plan 37

Letters of Welcome 3, 6, 20, 34 Exhibitor List 38 – 39

ISSCR Keynote Speaker 8 Exhibitor/Supporter Directory 41 – 57

Fourth Annual Anne McLaren Memorial Lecture 9 Detailed Program and Abstracts 59 – 93

Inaugural Ernest McCulloch Memorial Lecture 9 Wednesday, June 15 59 – 60

ISSCR Public Service Award 10 Thursday, June 16 61 – 71

ISSCR Outstanding Young Investigator Award 10 Friday, June 17 71 – 82

McEwen Centre Award for Innovation 11 Saturday, June 18 82 – 93

ISSCR Leadership and Staff 13, 24, 28, 32 Innovation Showcases 94 – 97

Detailed Schedule at a Glance 15 – 35 Poster Floor Plan 98 – 99

ISSCR Future Annual Meetings 16, 104 Author Index 101 – 103

Travel Award Winners 18, 22, 26, 30

What’s New for 2011 Meeting Orientation Area Need a quick overview of the ISSCR Annual Meeting? First time to the meeting or to the Metro Toronto Convention Centre? Stop by the ISSCR Orientation Area located on the 600 level near registration for a five-minute virtual tour, what’s new and what not to miss. Attend and be entered to win: • Two tickets to visit the CN Tower • Dinner for two at the Hard Rock Café

“Private Philanthropy and the Future of Stem Cell Research” luncheon panel Robert Klein, , Gordon Keller and Cheryl McEwen will demystify the process of finding and securing private funding. Friday, June 17, 11:30 – 1:30 p.m. See page 14 for details.

Two New ISSCR Awards Honoring Contributions to the Stem Cell Field Join us for the inaugural presentations of the ISSCR Public Service Award and the ISSCR McEwen Centre Award for Innovation, in addition to the third annual ISSCR Outstanding Young Investigator Award. See pages 11 for details about the recipients and presentation days and times.

Ernest McCulloch Memorial Lecture The ISSCR Board of Directors is pleased to establish the Ernest McCulloch Memorial Lecture to honor and celebrate his pioneering work. This inaugural lecture will be presented by John E. Dick during Plenary IV on Thursday. See page 9 for more information.

Lunch in the Exhibit Hall Now you don’t have to leave the convention center to grab something to eat and continue networking with your colleagues. Lunch is available for purchase in the Exhibit Hall (Concessions) on Thursday, Friday and Saturday. 7 ISSCR 9th Annual Meeting www.isscr.org

Don’t Miss these Featured Presentations

ISSCR Keynote Speaker Presidential Symposium: Stem and the Prospects for Regenerative Medicine: Past, Present and Future Wednesday, 1:00 p.m.

Robert S. Langer is the David H. Koch Institute Professor, the highest faculty member honor at the Massachusetts Institute of Technology. Dr. Langer has written more than 1,100 articles, with approximately 760 issued and pending patents worldwide. Dr. Langer’s patents have been licensed or sublicensed to more than 220 pharmaceutical, chemical, biotechnology and medical device companies. He is the most cited engineer in history.

He served as a member of the United States Food and Drug Administration’s SCIENCE Board, the FDA’s highest advisory board, from 1995 – 2002 and as its Chairman from 1999 – 2002.

Dr. Langer has received more than 180 major awards including the 2006 United States National Medal of Science; the Prize, considered the equivalent of the Nobel Prize for engineers and the 2008 Millennium Prize, the world’s largest technology prize. He is the also the only engineer to receive the Gairdner Foundation International Award; 72 recipients of this award have subsequently received a Nobel Prize. In 1989 Dr. Langer was elected to the Institute of Medicine of the National Academy of Sciences, and in 1992 he was elected to both the National Academy of Engineering and to the National Academy of Sciences. He is one of few people ever elected to all three United States National Academies and, at age 43, the youngest in history to ever receive this distinction.

Forbes Magazine (1999) and Bio World (1990) named Dr. Langer as one of the 25 most important individuals in biotechnology in the world. Discover Magazine (2002) named him as one of the 20 most important people in this area. Forbes Magazine (2002) selected Dr. Langer as one of the 15 innovators worldwide who will reinvent our future. Time Magazine and CNN (2001) named Dr. Langer as one of the 100 most important people in America and one of the 18 top people in science or medicine in America (America’s Best). Parade Magazine (2004) selected Dr. Langer as one of six “Heroes Whose Research May Save Your Life.” He received his bachelor’s degree from Cornell University in 1970 and his Sc.D. from the Massachusetts Institute of Technology in 1974, both in Chemical Engineering.

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Anne McLaren Memorial Lecturer Plenary VIII: Regulatory Networks of Stem Cells Saturday, 4:00 p.m.

Nicole M. Le Douarin is Honorary Professor at the College de France and Permanent Secretary at the Academie des Sciences in Paris. Her research career started under the sponsorship of Pr. Etienne Wolff who was the Director of the Institut d’Embryologie et de Tératologie expérimentales du Centre National de la Recherche Scientifique et du Collège de France in Paris. Her Doctorate in Sciences was an experimental study on digestive tract and liver development in the chick embryo. After her Do. Sci., Dr. Le Douarin was appointed as Assistant Professor at the University of Nantes where she devised a cell marking technique based on the construction of embryonic chimeras between two species of birds, the chick and the Japanese quail. This technique met with great success among embryologists because, in contrast to methods used to label cells in the embryo, it had two major advantages, stability and specificity. By combining this cell labelling technique with molecular tools, Dr. Le Douarin applied it to several embryonic systems in which she brought about a profound renewal of the state of the art. Her main contributions concern the development of the neural primordium and the hemopoietic and immune system. In 1975, she was invited to take over the Directorship of the Institut d’Embryologie du Collège de France et du CNRS. She was appointed as Professor at the Collège de France in the Chair of “Embryologie cellulaire et moléculaire” in 1988. She has been elected in 2001 as the “Secrétaire Perpétuelle” of the Académie des Sciences de l’Institut de France of which she was a member since 1982

She is a member of several academics and has received numerous awards including: the Edwin Conklin Award of The American Society for Developmental Biology (San Francisco) in 2005, the Ralph W. Gerard Prize of Neuroscience, Society of Neuroscience, USA in 2007 and the Prix d’Honneur de l’INSERM Paris in 2009. She is Grand Croix de l’Ordre National du Mérite and Grand Officier de la Légion d’Honneur.

Ernest McCulloch Memorial Lecture Plenary IV: Stem Cells and Cancer – Biology and Drug Development Thursday, 4:00 p.m.

John Dick, PhD, is a Senior Scientist at Princess Margaret Hospital, Professor in the Dept. of Molecular Genetics , and Director of the Cancer Stem Cell Program at the Ontario Institute for Cancer Research. Dr. Dick is world-renowned for his work in the development of normal and leukemia human stem cell assays based on transplantation of immune-deficient mice, which has transformed the study of stem cells. He led the group that identified leukemia initiating cells and later colon-cancer initiating cells, providing support for the hierarchical organization of some human cancers and the concept of cancer stem cells. Considered a leader in his field, he has won numerous major awards including the Dameshek and Thomas Prizes (ASH), Noble Prize (NCIC), Award (CIHR), Clowes Memorial Award (AACR), Clifford Prize for Cancer Research (Australia), and shared the Diamond Jubilee Award (NCIC) with Drs. and Ernest McCulloch.

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Join Us in Honoring the Recipients of the 2011 ISSCR Awards

ISSCR Public Service Award Presidential Symposium Wednesday, 1:00 p.m. The inaugural ISSCR Public Service Award will be awarded to Robert Klein, Chairman of the Governing Board of the California Institute for Regenerative Medicine (CIRM), for his outstanding contribution of public service to the field of stem cell research and regenerative medicine. Mr. Klein’s commitment to advancing medical research began when his younger son was diagnosed with juvenile diabetes in 2001. Through his vision as author and champion of the California Stem Cell Research and Cures Initiative (Proposition 71), Bob Klein secured for California long-term financial support for stem cell research. Through his leadership at CIRM, Bob developed a framework that fosters new and established talent, innovative science and clinical application. Klein’s advocacy for stable financial support for stem cell research on the international stage has been unprecedented. Robert Klein, Chairman, Independent Citizens’ Join us for this special award presentation which will take place during the Oversight Committee, Presidential Symposium on the afternoon of Wednesday, June 15, 2011. The California Institute for Regenerative Medicine (CIRM) Supported by past and present members of the International Society for Stem Cell Research Board of Directors

ISSCR Outstanding Young Investigator Award Plenary II: Totipotency and Germline Development Wednesday, 4:15 p.m.

The 2011 ISSCR Outstanding Young Investigator Award will be presented to Robert Blelloch, MD, PhD, Assistant Professor in the Departments of Urology; Obstetrics, Gynecology and Reproductive Sciences; and Pathology at UCSF; Peter R. Carroll, MD Endowed Chair and member of the Eli and Edythe Broad Center of Regeneration Medicine and Stem Cell Research, the Center for Reproductive Sciences, and the UCSF Helen Diller Family Comprehensive Cancer Center. Supported by the University of Pittsburgh, this award recognizes the exceptional achievements of an investigator in the early part of his or her independent career in stem cell research. The outstanding caliber of work achieved by this year’s nominees demonstrates the vital role that early-career investigators are playing in the rapid Robert Blelloch, MD, PhD progress being made in stem cell research.

Dr. Blelloch has clearly established a novel and independent line of research, providing profound insights Supported by the into the post-transcriptional regulation of early mammalian development. Specifically he has uncovered University of Pittsburgh important roles for microRNAs in regulating cell cycle of embryonic stem cells (ESCs) and a role for these same microRNAs in promoting induced pluripotency. He has also revealed novel classes of small RNAs in embryonic stem cells (ESCs) and the role of microRNAs in regulating the switch between self-renewal and differentiation. In addition he has established that microRNA function is globally suppressed in the oocyte/pre-implantation embryo.

Join us for the 2011 award presentation which will take place during Plenary Session II on the afternoon of Wednesday, June 15.

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Join Us in Honoring the Recipients of the 2011 ISSCR Awards

ISSCR McEwen Centre Award for Innovation 2011 Recipients: Kazutoshi Takahshi, PhD, and Shinya Yamanaka, MD, PhD Plenary VIII: Regulatory Networks of Stem Cells Saturday, 4:00 p.m.

The McEwen Centre Award for Innovation will be presented to Kazutoshi Takahashi, PhD, Lecturer, Center for iPS Cell Research and Application, Kyoto University and Shinya Yamanaka, MD, PhD, Director, Center for iPS Cell Research and Application, Kyoto University and Professor, Institute for Integrated Cell-Material Sciences, Kyoto University, in recognition of their paradigm-shifting work demonstrating the reprogramming of somatic cells using transcription factors that has resulted in a rapid development of novel tools and strategies for use in the pursuit of better understanding and treating disease. Supported by the McEwen Centre for Regenerative Medicine in Toronto, the award specifically acknowledges both junior and senior investigators, recognizing original thinking and ground breaking Kazutoshi Takahashi, PhD Shinya Yamanaka, MD, PhD research pertaining to stem cells or regenerative medicine that opens CiRA Kyoto University CiRA Kyoto University new avenues of exploration towards the understanding or treatment of human disease or affliction.

Join us for the 2011 award presentation which will take place during the Plenary Session VIII on the afternoon of Saturday, June 18.

Supported by

11 Cellular Programs and Reprogramming October 24–28, 2011 Dushu Lake Conference Center • Suzhou, China

Meeting Topics: Human Stem Cells Differentiation Mechanisms Cell Lineage Control of Pluripotency Cell Interactions Cell Therapies Molecular Control

Call for abstracts: Present your work along with these Deadline August 19, 2011 distinguished speakers: Keynotes: Elaine Fuchs and Gordon M. Keller Plenary sessions: Ling-Ling Chen, Linzhao Cheng, Kenneth Chien, Kevin A. D’Amour, Hongkui Deng, Tariq Enver, Fred Gage, Yukiko Gotoh, Anna-Katerina Hadjantonakis, Laura Johnston, V. Narry Kim, Mark Krasnow, Linheng Li, Haifan Lin, Hiromitsu Nakauchi, Jennifer Nichols, Hitoshi Niwa, Hideyuki Okano, Duanqing Pei, Wolf Reik, Deepak Srivastava, Lorenz Studer, Toshio Suda, , Marius Wernig, Joanna Wysocka, Yi Zhang

Conference Organizers Hongkui Deng, School of Life Sciences, Peking University, China; V. Narry Kim, Seoul National University, Korea; Ronald McKay, Lieber Institute for Brain Development, USA; Richard A. Young, Whitehead Institute/M.I.T., USA. Registration and abstract submission at www.csh-asia.org/Joint11.html 12 www.isscr.org Final Program

ISSCR Leadership Executive Committee President Elaine Fuchs, PhD The

President-Elect Fred H. Gage, PhD Salk Institute for Biological Studies

Vice President Shinya Yamanaka, MD, PhD Kyoto University Center for iPS Cell Research and Application (CiRA)

Clerk Sir Ian Wilmut, OBE, FRS, FRSE University of Edinburgh

Treasurer Sally Temple, PhD NY Neural Stem Cell Institute

Past President Irving L. Weissman, MD Stanford University School of Medicine

Board of Directors

Margaret Buckingham, DPhil Ihor R. Lemischka, PhD Derek J. van der Kooy, PhD Pasteur Institute Mount Sinai Medical Center University of Toronto Hans C. Clevers, MD, PhD Haifan Lin, PhD Amy Wagers, PhD Hubrecht Institute Yale University School of Medicine Joslin Diabetes Center Hongkui Deng, PhD Olle Lindvall, MD, PhD Ex Officio Peking University University of Lund George Q. Daley, MD, PhD Children’s Hospital Boston , PhD Daniel R. Marshak, PhD Lab BC Cancer Agency PerkinElmer Inc. Paul J. Simmons, PhD University of Texas Health Science Center Lawrence S.B. Goldstein, PhD Sean J. Morrison, PhD University of California, San Diego University of Michigan/HHMI Fiona M. Watt, DPhil CRUK Cambridge Research Institute Thomas Graf, PhD Christine L. Mummery, PhD Center for Genomic Regulation DYC Leiden University Medical Center Leonard I. Zon, MD Children’s Hospital Boston Markus Grompe, MD David T. Scadden, MD Oregon Health & Science University Harvard Stem Cell Institute Massachusetts General Hospital

ISSCR Global Advisory Council

Robert N. Klein - Chair George Q. Daley, MD, PhD Rob McEwen Mukesh Ambani Gordon Keller, PhD Charles Sabine , PhD Jim Yong Kim, MD, PhD Paul J. Simmons, PhD Gregory Bonfiglio, JD William Kridel, PhD Leonard I. Zon, MD Gerald Lokchung Chan, DSc Susan Lim, FRCS, PhD 13 ISSCR 9th Annual Meeting www.isscr.org

“Private Philanthropy and the Future of Stem Cell Research” Insights for building successful private funding relationships. Friday, June 17 11:30 a.m. – 1:30 p.m. InterContinental Toronto Centre

Learn how private philanthropy support is garnered, how it has shaped stem cell research and the impact it will continue to have on the field. Our panel of experts will demystify the process of finding and securing private funding. Institute directors and principal investigators will gain strategic insights from both sides of successful private funding relationships. Robert N. Klein, Susan L. Solomon, Moderator Panelist Moderator: Robert N. Klein, Chairman, Independent Citizens’ Oversight Committee, The California Institute for Regenerative Medicine (CIRM)

Panelists: • Gordon M. Keller, PhD, Director, McEwen Centre for Regenerative Medicine Gordon M. Keller, PhD, Cheryl McEwen, Panelist Panelist • Cheryl McEwen, Co-Founder, McEwen Centre for Regenerative Medicine and Vice-Chair, Toronto General & Western Hospital Foundation

• Susan L. Solomon, Chief Executive Officer and Co-Founder, The New York Stem Cell Foundation (NYSCF)

Luncheon registration and additional fee are required. If you haven’t pre-registered for this event, go to the Registration Desk to check availability.

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Detailed Schedule at a Glance

Tuesday, June 14 Room Abstract Page # 2:00 – 6:00 p.m. Meeting Orientation 600 Level, near Registration

Wednesday, June 15 7:30 a.m. – 5:00 p.m. Meeting Orientation 600 Level, near Registration 8:30 a.m. – 12:30 p.m. Industry Wednesday Symposia BD Biosciences 700 Level – 701 AB How Flow Cytometry in Combination with Optimized Cell Culture Environments Can Enable the Stem Cell Workflow — from Isolation and Expansion to Analysis and Production, from Basic Research to Large-Scale Cell Processing Lonza Walkersville, Inc. 700 Level – 718A Novel Stem Cell Tools: Applications in Research, Drug Discovery and Cell Therapy Thermo Scientific 700 Level – 718B Practical Applications of Stem Cells and Stem Cell Technologies 9:00 a.m. – 12:30 p.m. Focus Sessions ISSCR Ethics and Public Policy Committee 700 Level – 716B Ethics for Researchers at the Frontiers of Science— the Case of Stem Cell Research Mount Sinai School of Medicine 700 Level – 714A Governance/Management/Policies of Human ESC/iPSC Shared Resource Facilities Stem Cell Network 700 Level – 714B An Informed Society—How to Participate in Public Science Education and Why it Matters University Health Network - Toronto, Cell Stem Cell and the ISSCR 700 Level – 717 AB Scientific Legacy: a Symposium to Honor Ernest McCulloch 1:00 – 3:30 p.m. Presidential Symposium: Stem Cell Biology and the Prospects for 800 Level Regenerative Medicine: Past, Present and Future Plenary Halls F & G Supported by COSAT - Johnson & Johnson Chair: Elaine Fuchs 1:00 – 1:20 p.m. Welcomes & State of the Society Address, ISSCR President Elaine Fuchs 1:20 – 1:30 p.m. Presentation of the ISSCR Public Service Award to Robert Klein, Chairman, Independent Citizens’ Oversight Committee, The California Institute for Regenerative Medicine (CIRM) 1:30 – 2:00 p.m. Janet Rossant, Hospital for Sick Children, Canada 59 FROM TILL AND MCCULLOCH TO IPS CELLS — THE IMPORTANCE OF FUNCTIONAL ASSAYS 2:00 – 2:30 p.m. George Q. Daley, Children’s Hospital Boston, USA 59 DIRECTING AND REDIRECTING CELL FATES 2:30 – 3:00 p.m. Keynote Address: Robert S. Langer, Massachusetts Institute of Technology, USA 59 REGENERATIVE MEDICINE: TISSUE ENGINEERING, BIOMATERIALS AND CONTROLLED DRUG DELIVERY 3:00 – 3:30 p.m. Irving L. Weissman, Stanford University School of Medicine, USA 59 TITLE TBD 3:00 – 8:00 p.m. Exhibits Open 800 Level – Exhibit Halls D & E 3:30 – 4:00 p.m. All Posters Put On Display Exhibit Halls D & E 3:30 – 4:15 p.m. Refreshment Break Exhibit Halls D & E 4:00 – 8:00 p.m. Posters Open for Viewing Exhibit Halls D & E

15 10th Save Annual the Date Meeting June 13 – 16, 2012 Pacifico Yokohama • Yokohama, Japan

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Co-sponsored by

iPSCs stained with pluripotency marker, © Fate Therapeutics 16 Expression of Thy-1 antigen on MSC, © Weiwen Deng, M.D., Ph.D. www.isscr.org Final Program

Detailed Schedule at a Glance

Wednesday, June 15 (continued) Room Abstract Page # 4:15 – 6:30 p.m. Plenary II: 800 Level – Totipotency and Germline Development Plenary Halls F & G Chair: Haifan Lin 4:16 – 4:41 p.m. Ruth Lehmann, HHMI/Skirball Institute, USA PROTECTING THE GERMLINE THROUGH SILENCING 4:42 – 5:07 p.m. Azim Surani, Wellcome Trust & Cancer Research UK Gordon Institute, United Kingdom 59 THE RELATIONSHIP BETWEEN SPECIFICATION OF GERM CELLS AND REGULATION OF PLURIPOTENCY 5:08 – 5:33 p.m. Hans Schoeler, Max Plank Institute, Germany 60 INDUCTION OF PLURIPOTENCY IN GERMLINE AND SOMATIC CELLS 5:34 – 5:59 p.m. Shosei Yoshida, National Institute for Basic Biology, Japan 60 SPERM STEM CELLS IN THE MOUSE TESTIS 6:00 – 6:30 p.m. Presentation of the Outstanding Young Investigator Award 60 Robert Blelloch, University of California, San Francisco, USA MICRORNAS TO PATHWAYS IN STEM CELL FATE DECISIONS 6:30 – 8:00 p.m. Opening Reception 800 Level – Exhibit Halls D & E Supported by the McEwen Centre for Regenerative Medicine 8:00 – 9:30 p.m. Junior Investigator Career Panel Fairmont Royal York Supported by Stemgent Imperial Room Strategies for Success: Making the Most of Funding Opportunities

Thursday, June 16 8:30 – 9:30 a.m. Morning Coffee 800 Level – Plenary Hall Foyer 9:00 – 11:30 a.m. Plenary III: 800 Level – Plenary Halls F & G Tissue Stem Cell Origins Supported by Lieber Institute for Brain Development Chair: Margaret Buckingham 9:01 – 9:26 a.m. Marianne Bronner, California Institute of Technology, USA 61 SPECIFICATION AND MAINTENANCE OF NEURAL CREST CELL FATE 9:27 – 9:52 a.m. Shin-Ichi Nishikawa, RIKEN Center for Developmental Biology, Japan 61 DEVELOPMENTAL PROCESS OF HEMATOPOIETIC STEM CELL EXPLAINED 9:53 – 10:18 a.m. Michael A. Rudnicki, Ottawa Health Research Institute, Canada 61 WNT SIGNALLING AND THE REGULATION OF MUSCLE STEM CELL FUNCTION 10:19 – 10:44 a.m. Elaine Dzierzak, Erasmus Stem Cell Institute, Netherlands 61 ENDOTHELIAL ORIGINS OF HEMATOPOIETIC STEM CELLS 10:45 – 11:10 a.m. Hans C. Clevers, Hubrecht Institute, Netherlands 62 LGR5 STEM CELLS IN SELF-RENEWAL AND CANCER 11:10 – 11:25 a.m. Poster Teasers 11:00 a.m. – 8:00 p.m. Exhibits and Posters Open 800 Level – Exhibit Halls D & E 11:30 a.m. – 1:30 p.m. Lunch on your own Lunch available for purchase in the Exhibit Hall 11:45 a.m. – 1:15 p.m. Meet the Experts Lunch 700 Level – 713 AB & 715 AB (Pre-registration required) 11:45 a.m. – 12:15 p.m. Innovation Showcases Scottish Development International 700 Level – 718 AB 94 Innovation in Scotland: Novel Stem Cell Tools and Technologies Presenters: David C. Hay, Verna McErlane, Jo Mountford DVS Sciences Inc. 700 Level – 716 AB 94 Multiparametric Analysis with Mass Cytometry Presenters: Dmitry Bandura, Olga Ornatsky Corning Incorporated 700 Level – 714 AB 94 Use of a Novel Synthetic Surface for the Derivation, Proliferation and Differentiation of iPS Cells and their Progeny Presenter: Michael J. Young 17 The Stem Cell Network is a proud supporter of the 2011 ISSCR Travel Awards

Congratulations to the following recipients

Rajesh Alphonse Borhane Guezguez P. Joel Ross Peggy Assinck Lamis Hammoud Ryan Salewski Tzvi Aviv S M Mahmudul Hasan Mehdi Shafa Steffen Biechele Karin Hermans Andrey Shukalyuk Melanie Bilodeau Corinne Hoesli Shreya Shukla Jennifer Bruin Megan Hunt Sabrina Sicilia Ashley Calder Rosario Isasi Darek Sikorski Pearl Campbell Melanie Kardel Rita Silva Paul Cassar Shahryar Khattak Kim Snyder Aaron Cheung Véronique Lecault Vahab Soleimani Chun-Yi Chiang Poh Soo Lee Hannah Song Brian Cox Bernard Lo Brenna Swift Elizabeth Csaszar Mandy Lo Mark Ungrin Eric Deneault Anthony Mak Jie Ting Tina Wang Julia DiLabio Angela McDonald WeiJia Wang Andrea Ditadi Michael Milyavsky Amy Wong Ugljesa Djuric Emanuel Nazareth Akihiro Yamashita Shaalee Dworski Thu Minh Nguyen Gustavo Yannarelli Natalie Farra M. Cristina Nostro Jonathan Yeh David Fluri Kento Onishi Telford Yeung Simon Fortier Krishna Panchalingam Hang Yin Catherine Frelin Ben Paylor Yan (Mary) Zhang Marco Gallo Feodor Price Regan-Heng Zhang Tia Gareau Iran Rashedi Sylvie Gervier Shanti Rojas-Sutterlin

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Detailed Schedule at a Glance

Thursday, June 16 (continued) Room Abstract Page # 11:45 a.m. – 12:15 p.m. Innovation Showcases (continued) STEMCELL Technologies Inc. 700 Level – 701 AB 94 STEMdiffTM APELTM, a Defined Animal Product Free Medium for the Growth Factor Directed Differentiation of Pluripotent Stem Cells Presenter: Elizabeth Ng BD Biosciences 800 Level – 801 AB 94 Integrated Solutions for the Expansion and Analysis of Human Mesenchymal Stem Cells Presenter: Abel Hastings 12:30 – 1:00 p.m. Innovation Showcases ATCC 700 Level – 718 AB 95 iPS Cell Repository for Human Tissue and Disease Models Presenter: Will Rust Stemgent 700 Level – 716 AB 95 Reproducible Derivation of Integration-Free Human iPS Cells using mRNA Presenter: Brad Hamilton BD Biosciences 700 Level – 714 AB 95 Isolation of Single Cells from SPKLS Sub-populations by Flow Cytometry for Downstream Single Cell Gene Expression Profiling Presenter: Gil Reinin Union Biometrica Inc. 700 Level – 701 AB 94 Large Particle Flow Cytometry for Cells and Cell Clusters in Stem Cell Research Presenter: Rock Pulak Molecular Devices Inc. 800 Level – 801 AB 94 Induced Pluripotent Stem Cells for Research, Drug Screening and Toxicity Testing Presenter: Jayne Hesley 1:30 – 3:15 p.m. Concurrent Session I Track A – Human iPS and Embryonic Stem Cells 700 Level – 718 AB Supported by Development Co-Chairs: Konrad Hochedlinger and Duanqing Pei 1:32 – 1:57 p.m. Konrad Hochedlinger, HHMI Harvard Medical School, USA 62 SOX2, STEM CELLS AND CELLULAR REPROGRAMMING 1:59 – 2:14 p.m. Amy P. Wong, Hospital for Sick Children, Canada 62 ESTABLISHMENT OF CFTR-EXPRESSING EPITHELIAL FROM PLURIPOTENT STEM CELLS 2:16 – 2:31 p.m. Shannon M. Buckley, Howard Hughes Medical Institute/NYU School of Medicine, USA 63 THE UBIQUITIN PROTEASOME SYSTEM REGULATES SELF RENEWAL AND DIFFERENTIATION OF MOUSE EMBRYONIC STEM CELLS 2:33 – 2:48 p.m. Natalia Ivanova, Yale University, USA 63 NANOG-OCT4-SOX2 REGULATORY MODULE IN HUMAN EMBRYONIC STEM CELLS 2:50 – 3:15 p.m. Duanqing Pei, Guangzhou Institute of Biomedicine & Health CAS, Peoples Republic of China 63 REPROGRAMMING UNDER OPTIMIZED CONDITIONS REVEALS A SHORTENED ROUTE TO PLURIPOTENCY Track B – Stem Cells & Tissue Engineering 700 Level – 716 AB Supported by Ontario Stem Cell Initiative (OSCI) Co-Chairs: Peter W. Zandstra and Laura E. Niklason 1:32 – 1:57 p.m. Peter W. Zandstra, University of Toronto; 64 McEwen Centre for Regenerative Medicine, University Health Network, Canada TOWARDS AN INTEGRATED SUSPENSION-BASED PLURIPOTENT STEM CELL INDUCTION, EXPANSION AND DIRECTED DIFFERENTIATION PLATFORM 1:59 – 2:14 p.m. Todd C. McDevitt, Georgia Tech / Emory University, USA 64 ENGINEERING THE 3D MICROENVIRONMENT OF MULTI-CELLULAR PLURIPOTENT STEM CELL AGGREGATES FOR DIRECTED DIFFERENTIATION AND MORPHOGENESIS 2:16 – 2:31 p.m. Hidetatsu Otani, Osaka University Graduate School of Medicine, Japan 65 DIRECTED INDUCTION OF CHONDROGENIC CELLS FROM MURINE DERMAL FIBLOBLAST CULTURE WITHOUT GOING THROUGH A PLURIPOTENT STEM CELL STATE

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Letter from the Premier

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Detailed Schedule at a Glance

Thursday, June 16 (continued) Room Abstract Page # 1:30 – 3:15 p.m. Concurrent Session I (continued) Track B – Stem Cells & Tissue Engineering 700 Level – 716 AB 2:33 – 2:48 p.m. Donny Hanjava-Putra, Johns Hopkins University, USA 65 CONTROLLING MORPHOGENESIS OF ENDOTHELIAL PROGENITORS TO GENERATE FUNCTIONAL MICROVASCULATURE IN A SYNTHETIC MATRIX 2:50 – 3:15 p.m. Laura E. Niklason, Yale School of Engineering, USA 65 ENGINEERED LUNGS FROM BIO-SCAFFOLDS AND PULMONARY CELLS Track C – Neural Stem Cells 700 Level – 714 AB Supported by StemCells Inc. Co-Chairs: Sally Temple and Ronald D. McKay 1:32 – 1:57 p.m. Sally Temple, New York Neural Stem Cell Institute, USA 66 ENVIRONMENTAL FACTORS REGULATING NEURAL STEM CELL LINEAGE PROGRESSION 1:59 – 2:14 p.m. Alysson R. Muotri, University of California, San Diego, USA 66 SYNAPTIC DEFECTS IN HUMAN NEURONS DERIVED FROM AUTISM SPECTRUM DISORDERS PATIENTS 2:16 – 2:31 p.m. Tzvi Aviv, Hospital for Sick Children Toronto, Canada 66 COMPARATIVE FUNCTIONAL GENOMIC RNAI SCREEN OF EMBRYONIC AND NEOPLASTIC NEURAL STEM CELLS 2:33 – 2:48 p.m. Xiaoqun Wang, University of California, San Francisco, USA 66 PEERING INTO STEM CELLS IN LIVE BRAIN: A NEW SUBTYPE OF NEUROGENIC PROGENITOR IN MOUSE NEOCORTEX 2:50 – 3:15 p.m. Ronald D. McKay, Lieber Institute, USA 67 A TRANSCRIPTIONAL MECHANISM LINKING SOX2 AND PARAHOX GENES IN ADULT TISSUES Track D – Stem Cells and Regeneration 700 Level – 701 AB Supported by The Hospital for Sick Children, Research Institute Co-Chairs: Freda Miller and Elly Tanaka 1:32 – 1:57 p.m. Freda Miller, Hospital for Sick Children, University of Toronto, Canada 67 DERMAL STEM CELLS: FROM THE TO THE SPINAL CORD 1:59 – 2:14 p.m. Kosta Pajcini, Stanford University, USA 67 A NOVEL APPROACH TO REGENERATIVE MEDICINE: REVERSION OF THE POSTMITOTIC STATE BY TRANSIENT DUAL KNOCKDOWN OF TUMOR SUPPRESSORS 2:16 – 2:31 p.m. Takayuki Tanaka, Kyoto University, Japan 67 DISEASE MODELING WITH INDUCED PLURIPOTENT STEM CELLS REVEALS THE PATHOGENESIS OF SOMATIC MOSAICISM IN AN NLRP3-DRIVEN AUTOINFLAMMATORY SYNDROME 2:33 – 2:48 p.m. Salvador Aznar Benitah, Center for Genomic Research (CRG), Spain 68 REGULATION OF MOUSE EPIDERMAL STEM CELLS NICHE HETEROGENEITY BY A MOLECULAR CLOCK 2:50 – 3:15 p.m. Elly Tanaka, Max Planck Institute of Molecular Cell Biology and 68 Genetics, DFG Research Center for Regenerative Therapies, Technical University of Dresden, Germany APPLYING REGENERATIVE CONCEPTS FOR THE CNS FROM THE SALAMANDER TO MAMMALIAN NEURAL STEM CELLS Track E – Stem Cell Signaling 800 Level – 801 AB Co-Chairs: Emi Nishimura and Daniela Drummond-Barbosa 1:32 – 1:57 p.m. Emi Nishimura, Tokyo Medical and Dental University, Japan 68 FOLLICLE STEM CELL PROVIDE A FUNCTIONAL NICHE FOR MELANOCYTE STEM CELLS

21 Congratulations to the 2011 ISSCR Travel Award recipients Ece Akhan Akhan Linda Jansson Piriya Sasajala Adi Alajem Zi-Bing Jin Katja Schulze Brígida Almeida Junghyun Jo Bryce Seifert Lixia Bai Pakpoom Kheolamai S.W. Steven Shaw Anannya Banga Toshihiro Kobayashi Hiroko Shimada Ori Bar-Nur Pooja Kumar Maria Shutova Uri Ben-David Anujith Kumar Ng Shyh-Chang Jacob Blumenthal Elisabeth Larsen Karim Si-Tayeb Tal Bruck Aaron Levine Esther Son Stefan Brunner Guangnan Li Hathaitip Sritanaudomchai Shannon Buckley Zhe Li Ryohichi Sugimura Mi Cai Yuchun Liu Zeinab Taghizadeh Andre Catic Ilana Livyatan Jignesh Tailor Stuart Chambers Angelo Lombardo Naoya Takayama Fang-Pei Chang Dongdong Ma Hitoshi Takizawa Chien-Wen Chen Yo Mabuchi Yu-zhen Tan Yee Sook Cho Shin-ichi Mae Koji Tanabe Hong Seo Choi Mohammad Mandegar Daiki Tateyama Hyung Jun Choi Sirikul Manochantr Derk ten Berge Diana Coronado Giacomo Masserdotti Adrian Teo Victor Dayan Anna Mattout Eirini Trompouki Valdo Jose Dias da Silva Yves Maury Alicia Vaca Dasa Dolezalova Freyja McClenahan Yu-Chieh Wang Kalina Draganova Shai Melcer Xiaoqun Wang Marleen Eijkholt Giulia Meneghello Qingjie Wang Kim Erwes Miguel Mitne Neto James Wood Fernando Ezquer Sujata Mohanty Min-Zu Wu Marcelo Ezquer Morvarid Mohseni Yao-Ming Wu Tiago Faial Dafni Moschidou Lina Xiang Yiping Fan Laura Muertz Liangqi Xie Faranak Fattahi Kristopher Nazor Cong Xu Joji Fujisaki Yifan Ng Swathi Yadlapalli Mariana Garcia Anja Nitzsche Kohei Yamamizu Marcela Garita-Hernandez Aya Ogura Taiji Yamazoe Astrid Gillich Mariaestela Ortiz Hongyao Yu Renee Gordon Bruna Paulsen AzadehZahabi Yvonne Gruber Mica Gioia Petrighi Polidori Zhiyong Zhang Donny Hanjaya-Putra Ulrich Pfisterer Jianhong Zhu Nicholas Hannan Sarah Pringle Lili Zhu Garrett Heffner Stephanie Protze Limor Zwi-Dantsis Ahmed Hegab Li Qian Dominic Heng Tali Re’em Huimei Hong Paulo Henrique Rosado de Justin Ichida Castro Banu Iskender Ricardo Rossello 22 www.isscr.org Final Program

Detailed Schedule at a Glance

Thursday, June 16 (continued) Room Abstract Page # Track E – Stem Cell Signaling (continued) 800 Level – 801 AB 1:59 – 2:14 p.m. Eli R. Zunder, Stanford University, USA 69 HIGH CONTENT, SINGLE CELL ANALYSIS OF EMBRYONIC STEM CELL DIFFERENTIATION BY MASS CYTOMETRY 2:16 – 2:31 p.m. Joseph R. Klim, University of Wisconsin Madison, USA 69 TAILORED SYNTHETIC SURFACES TO CONTROL HUMAN PLURIPOTENT STEM CELL FATE 2:33 – 2:48 p.m. Anthony B. Mak, University of Toronto, Canada 69 THE CANCER STEM CELL MARKER CD133 IS REGULATED BY HISTONE DEACETYLASE 6 AND FUNCTIONS TO STABILIZE BETA-CATENIN IN WNT SIGNALING FOR SUPPRESSION OF CANCER CELL DIFFERENTIATION 2:50 – 3:15 p.m. Daniela Drummond-Barbosa, Johns Hopkins Bloomberg School of Public Health, USA 70 CONTROL OF STEM CELLS BY DIET AND SYSTEMIC FACTORS IN THE DROSOPHILA OVARY 3:15 – 4:00 p.m. Refreshment Break 800 Level – Exhibit Halls D & E 4:00 – 6:00 p.m. Plenary IV: 800 Level – Plenary Halls F & G Stem Cells and Cancer – Biology and Drug Development Chair: David T. Scadden 4:01 – 4:26 p.m. Thea D. Tlsty, University of California, San Francisco, USA 70 PHENOTYPIC PLASTICITY IN HUMAN SOMATIC CELLS 4:27 – 4:52 p.m. Pier Paolo Di Fiore, IFOM-IEO Campus, University of Milan, Italy 70 ENDOCYTOSIS STEM CELLS AND CANCER 4:53 – 5:18 p.m. Michael Clarke, Stanford University, USA 71 CLINICAL IMPLICATIONS OF EPIGENETIC REGULATORS OF SELF RENEWAL 5:19 – 5:54 p.m. The Ernest McCulloch Memorial Lecture 71 John E. Dick, University Health Network, Canada THE GENETIC DIVERSITY OF LEUKEMIA INITIATING CELLS OCCURS THROUGH A COMPLEX EVOLUTIONARY PROCESS 5:55 – 6:10 p.m. Poster Teasers 6:10 – 8:00 p.m. Poster Presentation/Exhibit Reception 800 Level – Exhibit Halls D & E 9:00 p.m. – 12:00 a.m. Junior Investigator Night Club Party Supported by Stemgent

Friday, June 17 8:30 – 9:30 a.m. Morning Coffee 800 Level – Plenary Hall Foyer 9:00 – 11:30 a.m. Plenary V: 800 Level – Plenary Halls F & G Reprogramming and Fate Conversion Supported by The New York Stem Cell Foundation (NYSCF) Chair: Gordon M. Keller 9:01 – 9:26 a.m. Shinya Yamanaka, Center for IPS Cell Research & Application, Japan 71 INDUCTION OF PLURIPOTENCY BY DEFINED FACTORS 9:27 – 9:52 a.m. John Gurdon, The Gurdon Institute, United Kingdom 72 THE DIRECT REPROGRAMMING OF SOMATIC CELL NUCLEI TO AN EGG OR OOCYTE PATTERN OF GENE EXPRESSION 9:53 – 10:18 a.m. Thomas Graf, Center for Genomic Regulation, CRG, Spain 72 TET2 PROMOTES C/EBPA-INDUCED PRE-B CELL TRANSDIFFERENTIATION BY DE-REPRESSING MACROPHAGE GENES 10:19 – 10:44 a.m. Andras Nagy, Mount Sinai Hospital, Canada 72 TRANSPOSON-MEDIATED REPROGRAMMING PROVIDES A POWERFUL TOOL FOR UNDERSTANDING SOMATIC CELL REPROGRAMMING TO PLURIPOTENCY 10:45 – 11:10 a.m. , Whitehead Institute for Biomedical Research, USA 73 STEM CELLS, PLURIPOTENCY AND NUCLEAR REPROGRAMMING 11:10 – 11:25 a.m. Poster Teasers 11:00 a.m. – 8:00 p.m. Exhibits and Posters Open 800 Level – Exhibit Halls D & E 23 ISSCR 9th Annual Meeting www.isscr.org

ISSCR Leadership

Annual Meeting Ethics and Public Policy Junior Investigators Public Education Committee Program Committee Committee Committee Rodney L. Rietze, PhD - Chair Margaret H. Baron, MD, PhD Haifan Lin, PhD – Chair Jan Helge Solbakk, MD, MA, Gerald De Haan, PhD - Chair Don Gibbons, PhD Kenneth R. Chien, MD, PhD PhD - Chair Hanna K.A. Mikkola, MD, Pamela J. Hines, PhD Linheng Li, PhD Angela Ballantyne, BSc, PhD PhD - Chair William Hoffman Daniel R. Marshak, PhD Timothy Caulfield, LLM, FRSC Cédric Blanpain, MD, PhD Suzanne Kadereit, PhD Ronald D. McKay, PhD Lawrence S.B. Goldstein, PhD Agnieszka Czechowicz, PhD M. William Lensch, PhD Shin-Ichi Nishikawa, MD, PhD Erica Haimes, BA, PhD Christos Gekas, PhD Angela C.H. McDonald, HBSC Janet Rossant, BA, PhD Insoo Hyun, PhD Garrett Heffner, PhD Richard A. Mollard, MBA, PhD Azim Surani, PhD Kazuto Kato, PhD Tara L. Huber, BA, PhD Sergio Pistoi, PhD Amy Wagers, PhD Jonathan Kimmelman, PhD Leanne Jones, PhD Priscilla E.M. Purnick, PhD Yukiko Yamashita, PhD Jason Scott Robert, PhD Hoang Nguyen, PhD Lisa Willemse Richard A. Young, PhD Christopher Thomas Scott Mariaestela Ortiz, PhD Leonard I. Zon, MD Loane Skene, LLB, LLM, LLD Nilay Thaker Elaine Fuchs, PhD - Ex Officio Jeremy Sugarman, MD, MPH, Publications Committee MA Legislative Educational Haifan Lin, PhD - Chair Audit Committee Patrick Taylor, JD Konrad Hochedlinger, PhD Giuseppe Testa, MD, PhD Initiative Kathrin Plath, PhD Carla Kim, PhD - Chair Prathap Tharyan, MD, George Q. Daley, MD, PhD - Toshio Suda, MD Margaret H. Baron, MD, PhD MRCPsych Chair Fiona Watt, DPhil Allen C. Eaves, MD, PhD, Lawrence S.B. Goldstein, Leonard I. Zon, MD FRCPC PhD - Chair Martin F. Pera, PhD Finance Committee Sean J. Morrison, PhD - Chair Sally Temple, PhD - Chair Web Site Advisory Committee R. Alta Charo, JD Gregory A. Bonfiglio, JD Jan Helge Solbakk, MD, MA, Hans-Willem E. Snoeck, MD, Awards Committee Hans C. Clevers, MD, PhD PhD PhD - Chair Fred H. Gage, PhD - Chair Andrew George Elefanty, MB, Alan O. Trounson, MSc, PhD Cédric Blanpain, MD, PhD Hans C. Clevers, MD, PhD BS, FRACP, PhD Sir Ian Wilmut, FRS, FRSE, OBE Fernando Camargo, PhD George Q. Daley, MD, PhD Danielle Guttman-Klein Shinya Yamanaka, MD, PhD Timothy Caulfield, LLM, FRSC John E. Dick, PhD Gordon M. Keller, PhD Elaine Fuchs, PhD – Paul J. Gadue, PhD, BS Margaret T. Fuller, PhD Robert N. Klein Ex Officio Member Fernando Pitossi, PhD Sir John B. Gurdon, DPhil, Sean J. Morrison, PhD Rodney L Rietze, PhD DSc, FRS Douglas A. Sipp Hiromitsu Nakauchi, MD, PhD Industry Committee Membership Committee Jan Helge Solbakk, MD, MA, Martin F. Pera, PhD - Chair PhD Jane S. Lebkowski, PhD - Chair David Bryder, PhD Fiona M. Watt, DPhil Clinical Translation Committee Ann Tsukamoto, PhD - Chair Sheng Ding, PhD David T. Scadden, MD - Chair Timothy Allsopp, BSc, PhD Toru Nakano, MD, PhD Alok Srivastava, MD, FRACP, Timothy Bertram, DVM, PhD Douglas A. Sipp FRCPA - Chair Ramkumar Mandalam, PhD Giulio Cossu, MD Chris Mason, MBBS, PhD, Marie Csete, MD, PhD FRCS Nominating Committee Michele De Luca, MD Sridaran Natesan, PhD Irving L. Weissman, MD - Chair Claude Gerstle, MD Kazuhiro Sakurada, PhD George Q. Daley, MD, PhD Armand Keating, MD John D. Sinden, PhD Elaine Fuchs, PhD Issei Komuro, MD, PhD Matthias Steger, MBA Fred H. Gage, PhD Olle Lindvall, MD, PhD Berta Strulovici, PhD Gordon M. Keller, PhD Maria Millan, MD Paul J. Simmons, PhD Xuetao Pei, PhD, MD Fiona M. Watt, DPhil Etienne Sokal, MD International Affairs Shinya Yamanaka, MD, PhD Deepak Srivastava, MD Nissim Benvenisty, MD, PhD - Leonard I. Zon, MD Anna Veiga, PhD Chair Gordon C. Weir, MD Ronald D. McKay, PhD - Chair Linzhao Cheng, PhD Douglas A. Melton, PhD Irving L. Weissman, MD Richard A. Young, PhD

24 www.isscr.org Final Program

Detailed Schedule at a Glance

Friday, June 17 (continued) Room Abstract Page # 11:30 a.m. – 1:30 p.m. Lunch on your own Lunch available for purchase in the Exhibit Hall 11:45 a.m. – 1:15 p.m. Meet the Experts Lunch 700 Level – 713 AB & 715 AB (Pre-registration required) 11:45 a.m. – 12:15 p.m. Innovation Showcases BD Biosciences 700 Level – 718 AB 96

Multiparameter Flow Cytometry and Bioimaging: Tools for Identifying Unique Immunophenotypes to Isolate and Analyze Sub-populations of Stem Cells and their Derivatives Presenter: Christian Carson Fluidigm 700 Level – 716 AB 96 Isolation of Mouse Hematopoietic Stem Cell Side Populations Using SPKLS and Post-Sort Confirmation Using Single-Cell Expression Presenters: Ken Livak and Alain Mir Miltenyi Biotec GmbH 700 Level – 714 AB 95 New Techniques to Control Pluripotent Stem Cell Differentiation Presenter: Andreas Bosio Beckman Coulter Inc. 700 Level – 701 AB 95 Multiplex Gene-Expression Assay for Human Induced Pluripotented Stem Cells (iPSCs) Using GexP Genetic Analysis System Presenter: Bee-Na Lee STEMCELL Technologies Inc. 800 Level – 801 AB 95 StemAdhereTM: A Defined and Entirely Human Substrate for the Culture of hESCs and hiPSCs Presenter: Stephen A. Duncan 12:30 – 1:00 p.m. Innovation Showcases BD Biosciences 700 Level – 718 AB 97 Multiparameter Flow Cytometry: Tools for Isolating Pluripotent Stem Cells and Analyzing Endoderm, Ectoderm and Mesoderm Lineages Presenter: Nil Emre Stemgent 700 Level – 716 AB 96 A Highly Efficient RNA Transfection Reagent for the Manipulation of Cell Fate Presenter: Kerry P. Mahon Miltenyi Biotec GmbH 700 Level – 714 AB 96 Integrated Cell-Processing Device for Automated Manufacturing of GMP-Compliant Stem Cell Products Presenter: Stefan Miltenyi Sigma Life Science 700 Level – 701 AB 96 A Leading Global Stem Cell Research and Preclinical R&D Partner Presenter: John Listello R&D Systems Inc. 800 Level – 801 AB 96 Ex Vivo Expansion of Stem/Progenitor Cells Using Defined, Serum-Free Systems Presenter: Jessie H.-T. Ni 1:30 – 3:15 p.m. Concurrent Session II Track A – Hematopoietic Stem Cells 700 Level – 718 AB Co-Chairs: Tannishtha Reya and Andreas Trumpp 1:32 – 1:57 p.m. Tannishtha Reya, University of California, San Diego, USA 73 DEVELOPMENTAL MECHANISMS IN STEM CELLS AND CANCER 1:59 – 2:14 p.m. Momoko Yoshimoto, Indiana University School of Medicine, USA 73 MULTI-POTENT HEMATOPOIETIC PROGENITORS ARISE AT THE EXTRA-EMBRYONIC YOLK SAC PRIOR TO HEMATOPOIETIC STEM CELL EMERGENCE IN THE MOUSE EMBRYO 2:16 – 2:31 p.m. Trista E. North, Harvard Medical School, USA 74 METABOLISM-INDUCED REACTIVE OXYGEN SPECIES (ROS) AND HIF1α STIMULATION CONTROL THE INDUCTION AND EXPANSION OF HEMATOPOIETIC STEM CELLS 2:33 – 2:48 p.m. Rong Lu, Stanford University, USA 74 TRACKING MURINE HEMATOPOIETIC STEM CELL DIFFERENTIATION IN VIVO WITH SINGLE CELL PRECISION: NEW INSIGHTS INTO THE CLONAL SUCCESSION VERSUS CLONAL STABILITY DEBATE 25 The Australian Stem Cell Centre is a proud supporter of the 2011 ISSCR Travel Awards Congratulations to the following recipients Yen-Shun Chen Tracy Heng Danika Khong Juan Carlos Polanco Chew-Li Soh Nilay Thakar Charlotte Yap Qing Cissy Yu

26 www.isscr.org Final Program

Detailed Schedule at a Glance

Friday, June 17 (continued) Room Abstract Page # 1:30 – 3:15 p.m. Concurrent Session II (continued) 700 Level – 718 AB 2:50 – 3:15 p.m. Andreas Trumpp, Deutsches Krebsforschungszentrum (DKFZ), Germany 74 STRESS INDUCED ACTIVATION OF HSCS Track B – Stem Cells in Non-Mammalian Models 700 Level – 716 AB Co-Chairs: Debbie Yelon and Phillip Newmark 1:32 – 1:57 p.m. Debbie Yelon, University of California, San Diego, USA 75 DEFINITION AND DYNAMICS OF THE CARDIAC PROGENITOR POOL IN ZEBRAFISH 1:59 – 2:14 p.m. Ricardo A. Rossello, Duke University, HHMI, USA 75 MAMMALIAN GENES INDUCE IPS-LIKE CELLS IN NON-MAMMALIAN SPECIES 2:16 – 2:31 p.m. Dongdong Ma, Brigham and Women’s Hospital, USA 75 THE EPIGENETIC REGULATION OF RENAL STEM CELL NUMBER IN ZEBRAFISH 2:33 – 2:48 p.m. Owen J. Tamplin, Children’s Hospital Boston, USA 76 TRAFFICKING OF ZEBRAFISH HEMATOPOIETIC STEM CELLS DURING EMBRYONIC DEVELOPMENT 2:50 – 3:15 p.m. Phillip Newmark, University of Illinois at Urbana-Champaign, USA 76 INTESTINAL RENEWAL AND REGENERATION IN THE PLANARIAN SCHMIDTEA MEDITERRANEA Track C – Stem Cell Asymmetry 700 Level – 714 AB Co-Chairs: Juergen Knoblich and Wieland Huttner 1:32 – 1:57 p.m. Juergen Knoblich, IMBA - Institute of Molecular Biotechnology, Austria 77 ASYMMETRIC CELL DIVISION AND TUMORIGENESIS IN NEURAL STEM CELL LINEAGES 1:59 – 2:14 p.m. Ryohichi Sugimura, Stowers Institute, USA 77 A ROLE OF NON-CANONICAL WNT SIGNALING IN MAINTAINING HEMATOPOIETIC STEM CELLS 2:16 – 2:31 p.m. Scott E. Williams, The Rockefeller University, USA 77 ASYMMETRIC CELL DIVISIONS PROMOTE NOTCH-DEPENDENT EPIDERMAL DIFFERENTIATION 2:33 – 2:48 p.m. Swathi Yadlapalli, University of Michigan, USA 77 CHROMOSOME STRAND SEGREGATION DURING DROSOPHILA MALE GERMLINE STEM CELL DIVISION 2:50 – 3:15 p.m. Wieland Huttner, Max Planck Institute of Molecular Cell Biology and Genetics, Germany 78 NEURAL STEM AND PROGENITOR CELLS – A CELL BIOLOGICAL AND EVOLUTIONARY PERSPECTIVE Track D – Epithelial Stem Cells 700 Level – 701 AB Co-Chairs: Fiona M. Watt and 1:32 – 1:57 p.m. Fiona M. Watt, CRUK Cambridge Research Institute, 78 Cambridge University, United Kingdom RECIPROCAL SIGNALLING BETWEEN EPIDERMAL STEM CELLS AND CELLS IN THE NICHE 1:59 – 2:14 p.m. Sarah Kozar, Cambridge Research Institute, United Kingdom 78 MITOTIC CHRONOLOGIES IN MURINE GUT PROGENITORS 2:16 – 2:31 p.m. Lixia Bai, Fred Hutchinson Cancer Research, USA 78 IDENTIFICATION OF MAMMARY CANCER STEM CELLS USING TG11.5KB-GFP MICE 2:33 – 2:48 p.m. Morvarid Mohseni, Harvard University/Children’s Hospital 79 YAP1 ACTS DOWNSTREAM OF ALPHA-CATENIN TO CONTROL EPIDERMAL PROLIFERATION 2:50 – 3:15 p.m. Valerie Horsley, Yale Stem Cell Center, USA 79 ADIPOCYTES REGULATE THE SKIN STEM CELL NICHE Track E – Endodermal Stem Cells 800 Level – 801 AB Co-Chairs: Carla Kim and Hans-Willem E. Snoeck 1:32 – 1:57 p.m. Carla Kim, Children’s Hospital Boston & Harvard Medical School, USA 79 STEM CELL APPROACHES TO DISSECT LUNG CANCER BIOLOGY 1:59 – 2:14 p.m. Ahmed E. Hegab, Mattel Children’s Hospital UCLA, USA 80 A NOVEL STEM/PROGENITOR CELL POPULATION FROM MURINE TRACHEAL SUBMUCOSAL GLAND DUCTS WITH MULTIPOTENT REGENERATIVE POTENTIAL 27 ISSCR 9th Annual Meeting www.isscr.org

ISSCR Abstract Review Committee

Gregor B. Adams, PhD Valerie Horsley, PhD Duan-qing Pei, PhD Suneet Agarwal, MD, PHD Akitsu Hotta, PhD Mirza Peljto, PhD Juan Carlos Izpisua Belmonte, PhD Guang Hu, PhD John M. Perry, PhD Walter Birchmeier, PhD Wieland Huttner, PhD Kathrin Plath, PhD Cedric Blanpain, MD, PhD Insoo Hyun, PhD Shahin Rafii, MD Maria A. Blasco, PhD Stefan Irion, MD Anjana Rao, PhD Helen M. Blau, PhD Natalia B. Ivanova, PhD Donald Eugene Redmond, Jr, MD Catherine Bollard, MBChB, MD Kim B. Jensen, PhD Tannishtha Reya, PhD Tewis Bouwmeester, PhD Ying Jin, MD PhD Paul Riley, PhD Laurie Boyer, PhD Naihe Jing, PhD Jerome Ritz, MD Christopher K. Breuer, MD Kazuto Kato, PhD Jason Scott Robert, PhD Marianne Bronner, PhD Steve Kattman, PhD Matthew S. Rodeheffer, PhD Margaret Buckingham, DPhil Armand Keating, MD Derrick J. Rossi, PhD Michael Buszczak, PhD Ali Khademhosseini, PhD Lee Rubin, PhD Fernando Camargo, PhD Carla Kim, PhD Hidetoshi Sakurai, MD, PhD Timothy Caulfield, LLM Jonathan Kimmelman, PhD David V. Schaffer, PhD Ling-Ling Chen, PhD Juergen A. Knoblich, PhD Timm T. Schroeder, PhD Linzhao Cheng, PhD Mark A. Krasnow, MD, PhD Christopher Scott, MA Susana Chuva de Sousa Lopes, PhD Diane S. Krause, MD, PhD Toshiharu Shinoka, MD, PhD Hans C. Clevers, MD, PhD Kelvin Lam, Ph.D. Jan Helge Solbakk, MD, MA, PhD Peter Coffey, BSc, DPhil David Michael Langenau, PhD Hongjun Song, PhD Chad Cowan, PhD Jonas Larsson, MD, PhD Didier Y. Stainier, PhD Marie Csete, MD, PhD Terry Lechler, PhD Yi Sun, PhD Xing Dai, PhD Richard T. Lee, MD Eugene Scott Swenson, MD, PhD Gerald De Haan, PhD Linheng Li, PhD Elly Tanaka, PhD Jill L. De Jong, MD, PhD Jinsong Li, PhD Sally Temple, PhD Michele De Luca, MD William Lowry, PhD Giuseppe Testa, MD, PhD Robert J. Deans, PhD Jun Lu, PhD David Traver, PhD Sheng Ding, PhD John W. Ludlow, PhD Linda van Laake, PhD Dennis E. Discher, PhD Maria Carolina Marchetto, PhD Juan-Jose Ventura, PhD Fiona Doetsch, PhD Erika L. Matunis, PhD Zhong Wang, PhD Ibrahim Domian, MD, PhD Todd McDevitt, PhD Marius Wernig, MD, PhD Massimo Dominici, MD Brad Merrill, PhD Holger Willenbring, MD, PhD Daniela Drummond-Barbosa, PhD Hanna K A Mikkola, MD, PhD Carl P. Wonders, PhD Ira J. Fox, MD Freda Miller, PhD Sean M. Wu, MD, PhD Margaret T. Fuller, PhD Christine L. Mummery, PhD Andrew Xiao, PhD Christos Gekas, PhD Masato Nakagawa, PhD Ting Xie, PhD Adam Giangreco, PhD Atsushi Nakano, MD, PhD Ren-He Xu, MD, PhD Wolfram Goessling, MD, PhD Garry A. Neil, MD Yukiko Yamashita, PhD Magdalena Goetz, PhD Phillip A. Newmark, PhD Qin Yan, PhD Cayetano Gonzalez, PhD Hoang Nguyen, PhD Debbie Yelon, PhD Marie Jose Goumans, PhD Laura E. Niklason, MD, PhD Yoshinori Yoshida, MD, PhD Thomas Graf, PhD Emi Nishimura, MD, PhD Peter W. Zandstra, PhD Valentina Greco, PhD Trista E. North, PhD Yi Zeng, PhD Richard Gregory, PhD Steve K.W. Oh, PhD Kang Zhang, MD, PhD Markus Grompe, MD Benjamin Ohlstein, MD, PhD Yi Zhang, PhD Jacob Hanna, MD, PhD Sean Palecek, PhD Xinyu Zhao, PhD Peiman Hematti, MD In-hyun Park, PhD Qiao Zhou, PhD Konrad Hochedlinger, PhD Robert Passier, PhD Leonard I. Zon, MD Kristen Hope, PhD Bret Pearson, PhD

28 www.isscr.org Final Program

Detailed Schedule at a Glance

Friday, June 17 (continued) Room Abstract Page # Track E – Endodermal Stem Cells (continued) 800 Level – 801 AB 2:16 – 2:31 p.m. M. Cristina Nostro, McEwen Centre for Regenerative Medicine, Canada 80 TGFβ FAMILY MEMBERS AND WNT SIGNALING REGULATE PANCREATIC SPECIFICATION OF HUMAN PLURIPOTENT STEM CELLS 2:33 – 2:48 p.m. Valerie Gouon-Evans, Mount Sinai School of Medicine, USA 81 THE ENDOTHELIAL CELL NICHE COORDINATES HEPATIC SPECIFICATION THROUGH DUAL REPRESSION OF WNT AND NOTCH SIGNALING 2:50 – 3:15 p.m. Hans-Willem E. Snoeck, Mount Sinai School of Medicine, USA 81 GENERATION OF ANTERIOR FOREGUT AND ITS DERIVATIVES FROM HUMAN PLURIPOTENT CELLS 3:15 – 4:00 p.m. Refreshment Break 800 Level – Exhibit Halls D & E 4:00 – 6:00 p.m. Plenary VI: 800 Level – Plenary Halls F & G Stem Cell Metabolism and Aging Chair: Connie Eaves 4:01 – 4:26 p.m. Sean J. Morrison, HHMI and University of Michigan Center for Stem Cell Biology, USA 81 SOX17 EXPRESSION CONFERS SELF-RENEWAL POTENTIAL AND FETAL STEM CELL CHARACTERISTICS UPON ADULT HEMATOPOIETIC PROGENITORS 4:27 – 4:52 p.m. Irina Conboy, University of California, Berkeley, USA 82 MODIFYING REGENERATIVE POTENTIAL AND CELL FATE WITHIN MYOGENIC LINEAGE 4:53 – 5:18 p.m. Margaret A. Goodell, Baylor College of Medicine, USA 82 EPIGENETIC REGULATION OF HEMATOPOIETIC STEM CELLS 5:19 – 5:44 p.m. Amy Wagers, Harvard University and Joslin Diabetes Center, USA 82 MODULATORS OF STEM CELL REGENERATIVE FUNCTION IN SKELETAL MUSCLE 5:45 – 6:00 p.m. Poster Teasers 6:00 – 8:00 p.m. Poster Presentation/Exhibit Reception 800 Level – Exhibit Halls D & E

Saturday, June 18 8:30 – 9:30 a.m. Morning Coffee 800 Level – Plenary Hall Foyer 9:00 – 11:30 a.m. Plenary VII: 800 Level – Plenary Halls F & G Therapeutic Approaches to Stem Cells Supported by Pfizer Neusentis and our Canadian colleagues Chair: Daniel R. Marshak 9:01 – 9:26 a.m. Leonard I. Zon, Children’s Hospital and Dana-Farber Cancer Institute, 82 HHMI, Harvard Stem Cell Institute, Harvard Medical School, USA STIMULATION OF THE WNT-PGE2 PATHWAY TO IMPROVE HSC SELF-RENEWAL 9:27 – 9:52 a.m. Kenneth R. Chien, Massachusetts General Hospital, USA 83 DRIVING HEART PROGENITOR FATE IN VIVO WITH MODIFIED RNA 9:53 – 10:18 a.m. Christopher K. Breuer, Yale University School of Medicine, USA 83 THE DEVELOPMENT AND TRANSLATION OF THE TISSUE ENGINEERED VASCULAR GRAFT: FROM THE BENCH TO THE BEDSIDE AND BACK AGAIN 10:19 – 10:44 a.m. Michele De Luca, University of Modena and Reggio Emilia, Italy 83 LIMBAL STEM-CELL THERAPY AND LONG-TERM CORNEAL REGENERATION 10:45 – 11:10 Sheng Ding, Gladstone Institute of Cardiovascular Disease, USA 84 A CHEMICAL APPROACH TO CONTROLLING CELL FATE 11:11 – 11:26 a.m. Charles Sabine, Patient Advocate, UK 11:00 a.m. – 4:00 p.m. Exhibits and Posters Open 800 Level – Exhibit Halls D & E 11:30 a.m. – 1:30 p.m. Lunch on your own Lunch available for purchase in the Exhibit Hall

29 ISSCR 9th Annual Meeting www.isscr.org

BD Biosciences is a proud supporter of the 2011 ISSCR Travel Awards

Congratulations to the following recipients

Jerome Artus Daniela Bueno Na-Yu Chia Brad Dykstra Sanae Hamanaka Chia-Ling Hsieh Peggy Janich Joseph Klim Rong Lu Daniel Ortmann Sharon Paige Tea Soon Park Nils Pfaff Enrique Salero Gen Shinoda Eszter Varga

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Detailed Schedule at a Glance

Saturday, June 18 (continued) Room Abstract Page # 11:45 a.m. – 12:15 p.m. Innovation Showcases Biological Industries Ltd 700 Level – 716 AB 97 NutriStem™ hESC/iPSC XF- The First Universal Low Xeno Free Media to Culture Pluripotent Stem Cells Presenter: Michal Amit Life Technologies 700 Level – 714 AB 97 Xeno-Free Derivation and Maintenance of Pluripotent Cell Lines Presenter: Jordan E. Pomeroy EMD Millipore 700 Level – 701 AB 97 Advances in Reprogramming Efficiency and Culture of iPS Cells Presenter: Vi Chu 12:30 – 1:00 p.m. Innovation Showcases Life Technologies 700 Level – 714 AB 97 Micro RNA and Epigenetic Regulation of Neural Stem Cell Differentiation Presenter: Ron Hart 1:30 – 3:15 p.m. Concurrent Session III Track A – Cell Therapy 700 Level – 718 AB Supported by F. Hoffman-La Roche Ltd. Co-Chairs: Christine Mummery and Peter Coffey 1:32 – 1:57 p.m. Christine L. Mummery, Leiden University Medical Center, Netherlands 84 CARDIOMYOCYTES FROM PLURIPOTENT STEM CELLS IN GENETIC CARDIAC DISEASE AND DRUG SAFETY PHARMACOLOGY 1:59 – 2:14 p.m. Tea Soon Park, Johns Hopkins School of Medicine, USA 84 NONVIRAL CORD BLOOD-DERIVED IPSC POSSESS AUGMENTED CAPACITIES FOR GENERATING FUNCTIONAL AND ENGRAFTABLE ANGIOBLASTS IN A RETINAL ISCHEMIA MODEL 2:16 – 2:31 p.m. Armand Keating, University Health Network, Canada 85 MESENCHYMAL STROMAL CELLS MEDIATE THE SWITCH TO ALTERNATIVELY ACTIVATED MONOCYTES/MACROPHAGES AFTER ACUTE MYOCARDIAL INFARCTION 2:33 – 2:48 p.m. Chien-Wen Chen, University of Pittsburgh, USA 85 THE PARACRINE MECHANISM OF ANTI-FIBROTIC AND ANTI-INFLAMMATORY EFFECTS OF HUMAN PERIVASCULAR STEM CELLS 2:50 – 3:15 p.m. Peter Coffey, University College London, UK 86 STEMMING VISION LOSS USING STEM CELLS - SEEING IS BELIEVING Track B: Small Molecule Approach to Stem Cells 700 Level – 716 AB Supported by Centre for Commercialization of Regenerative Medicine Co-Chairs: Tewis Bouwmeester and Alan Ezekowitz 1:32 – 1:57 p.m. Tewis Bouwmeester, NIBR – Novartis Institutes for Biomedical Research, 86 Novartis Pharma AG, Switzerland TOWARDS IDENTIFYING FACTORS THAT CAN AID IN TISSUE REGENERATION 1:59 – 2:14 p.m. Justin K. Ichida, Harvard Stem Cell Institute, USA 86 MECHANISTIC INSIGHTS INTO DEFINED-FACTOR REPROGRAMMING USING SMALL MOLECULES 2:16 – 2:31 p.m. Garrett C. Heffner, Children’s Hospital Boston, USA 87 SOLUBLE FACTORS SPECIFY THE GENERATION OF HEMATOPOIETIC PROGENITORS WITH MULTILINEAGE ENGRAFTMENT POTENTIAL IN ADULT RECIPIENT MICE FROM MOUSE EMBRYONIC STEM CELLS 2:33 – 2:48 p.m. Gabsang Lee, Memorial Sloan-Kettering Hospital, USA 87 DISCOVERY OF POTENTIAL THERAPEUTIC COMPOUNDS FOR FAMILIAL DYSAUTONOMIA USING PATIENT-SPECIFIC AND SYMPTOM-RELEVANT IPSC DERIVED NEURAL CREST PRECURSORS 2:50 – 3:15 p.m. Alan Ezekowitz, Merck Research Laboratories, USA 87 STEM CELL THERAPEUTICS: WHEN WILL PROMISE BECOME PROFITS?

31 ISSCR 9th Annual Meeting www.isscr.org

ISSCR Leadership

ISSCR Staff Nancy Witty Jenny Korshak Jill Hronek Executive Director Member Services Administrator Communications Director Heather Rooke, PhD Jim Donovan Meagan Comerford Science Director Conference Director Communications Manager Michael Hagedorn Christine Meehan Lisa Kamen Administrative Director Conference Administrator Communications Manager Glori Rosenson Madhuri Carson William Chandler Committees & Outreach Manager Support & Exhibits Manager Accountant Carl Wonders, PhD Shannon Baily Greg Schultz Scientific Affairs Manager Support and Exhibits Administrator Managing Partner

About the International Society for Stem Cell Research

The ISSCR is the only global trans-disciplinary science-based organization dedicated to stem cell research. The ISSCR Annual Meeting has become the premier international forum for the presentation of stem cell science. Founded in 2002, the society brings together investigators researching stem cells in many different organ systems and models. The ISSCR seeks to: • Promote and foster the exchange and dissemination of information and ideas relating to stem cells • Encourage the general field of research involving stem cells • Promote professional and public education in the areas of stem cell research and applications

ISSCR membership is open to stem cell and regenerative medicine investigators and clinicians in all environments including universities, medical schools, government agencies, for-profit and not-for-profit research laboratories and industries worldwide. ISSCR membership has grown to more than 3,800 in just nine years and includes the world’s leading stem cell scientists.

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Detailed Schedule at a Glance

Saturday, June 18 (continued) Room Abstract Page # Track C: Genomic Integrity 700 Level – 714 AB Co-Chairs: Maria Blasco and Cedric Blanpain 1:32 – 1:57 p.m. Maria Blasco, CNIO (Spanish National Cancer Research Centre), Spain 87 TELOMERE DYNAMICS AT THE INNER CELL MASS AND EMBRYONIC STEM (ES) CELLS INDICATE A LINK BETWEEN TELOMERE BIOLOGY AND PLURIPOTENCY 1:59 – 2:14 p.m. Athurva Gore, University of California, San Diego, USA 88 FUNCTIONAL CONSEQUENCES OF SOMATIC MUTATIONS IN HUMAN INDUCED PLURIPOTENT STEM CELLS 2:16 – 2:31 p.m. Uri Ben-David, The Hebrew University, Israel 88 GENE EXPRESSION PATTERNS REVEAL NOVEL TISSUE-SPECIFIC CHROMOSOMAL ABERRATIONS IN HUMAN PLURIPOTENT AND MULTIPOTENT STEM CELLS 2:33 – 2:48 p.m. Michael Milyavsky, Ontario Institute for Cancer Research, University Health Network, Canada 88 GENOME-WIDE FUNCTIONAL SCREEN FOR REGULATORS OF DNA DAMAGE RESPONSE IN HUMAN HEMATOPOIETIC STEM CELLS 2:50 – 3:15 p.m. Cedric Blanpain, IRIBHM, Université Libre de Bruxelles, Belgium 88 HAIR FOLLICLE STEM CELLS USE DIFFERENT MECHANISMS TO MEDIATE GENOME MAINTENANCE DEPENDING ON THEIR STAGES OF ONTOGENY Track D: Epigenetic Programming of Stem Cells 700 Level – 701 AB Co-Chairs: Yi Zhang and Anjana Rao 1:32 – 1:57 p.m. Yi Zhang, University of North Carolina, USA 89 ROLE OF TET IN DNA METHYLATION AND EMBRYONIC STEM CELL SELF-RENEWAL 1:59 – 2:14 p.m. Andrew Xiao, Yale School of Medicine, USA 89 HISTONE VARIANT PROTEIN H2A.X PLAYS A NOVEL ROLE IN STEM CELLS 2:16 – 2:31 p.m. Kevin Huang, University of California, Los Angeles, USA 89 THE ROLE OF DNA METHYLATION IN REGULATING TRANSCRIPTOME IN MOUSE EMBRYONIC STEM CELLS 2:33 – 2:48 p.m. Alessandra Giorgetti, CMRB, Spain 90 CORD BLOOD-DERIVED NEURONS BY ECTOPIC EXPRESSION OF SOX2 AND CMYC 2:50 – 3:15 p.m. Suneet Agarwal, Children’s Hospital Boston, USA 90 ROLE OF TET PROTEINS IN MOUSE EMBRYONIC STEM CELLS AND MYELOID TUMORIGENESIS Track E – Cardiac & Muscle Stem Cells 800 Level – 801 AB Co-Chairs: Mark A. Krasnow and Didier Y. Stainier 1:32 – 1:57 p.m. Mark A. Krasnow, HHMI, Stanford University, USA 90 RADIAL CONSTRUCTION OF AN ARTERIAL WALL 1:59 – 2:14 p.m. Li Qian, Gladstone Institutes, USA 91 IN VIVO REPROGRAMMING OF MURINE CARDIAC FIBROBLASTS INTO CARDIOMYOCYTES 2:16 – 2:31 p.m. Foteini Mourkioti, Stanford University, USA 91 ADULT STEM CELL EXHAUSTION DUE TO SHORT TELOMERES IN CHRONICALLY DAMAGED DYSTROPHIN-DEFICIENT MDX/MTR MICE MIMICS HUMAN DUCHENNE MUSCULAR DYSTROPHY 2:33 – 2:48 p.m. Malte Tiburcy, Georg-August-University Goettingen, Germany 91 ENGINEERED SKELETAL MUSCLE CONTAINS FUNCTIONAL SATELLITE CELL NICHES CAPABLE OF MUSCLE REGENERATION IN VITRO 2:50 – 3:15 p.m. Didier Y. Stainier, University of California, San Francisco, USA 92 PROGENITOR CELLS IN ZEBRAFISH ORGANOGENESIS 3:15 – 4:00 p.m. Refreshment Break 800 Level – Exhibit Halls D & E 4:00 p.m. Posters Dismantle

33 ISSCR 9th Annual Meeting www.isscr.org

Letter from the Mayor

34 www.isscr.org Final Program

Detailed Schedule at a Glance

Saturday, June 18 (continued) Room Abstract Page # 4:00 – 4:25 p.m. ISSCR Business Meeting 800 Level – Plenary Halls F & G ISSCR Executive Director Report Nancy Witty ISSCR Treasurer Report Sally Temple, New York Neural Stem Cell Institute, USA ISSCR Membership Q & A ISSCR Executive Committee President-Elect’s Address Fred H. Gage, Salk Institute for Biological Studies, USA 4:25 – 6:40 p.m. Plenary VIII: 800 Level – Plenary Halls F & G Regulatory Networks of Stem Cells Chair: Fred H. Gage 4:25 – 4:40 p.m. Presentation of the McEwen Centre Award for Innovation Recipients: Shinya Yamanaka and Kazutoshi Takahashi 4:41 – 5:06 p.m. Richard A. Young, Whitehead Institute for Biomed Research, USA 92 TRANSCRIPTIONAL CONTROL OF EMBRYONIC STEM CELLS 5:07 – 5:32 p.m. Stuart H. Orkin, Harvard Medical School, Dana-Farber Cancer Institute, USA 92 REGULATORY NETWORKS IN STEM CELLS AND CANCER 5:33 – 5:58 p.m. Judy Lieberman, The CBR Institute for Biomedical Research, USA 92 A GENOME-WIDE SIRNA SCREEN IDENTIFIES SELECTIVE INHIBITORS OF BASAL-LIKE BREAST TUMOR-INITIATING CELLS 5:59 – 6:34 p.m. Anne McLaren Memorial Lecture 93 Nicole M. Le Douarin, Academie Des Sciences, France THE NEURAL CREST, A PLURIPOTENT STRUCTURE OF THE VERTEBRATE EMBRYO 6:35 – 6:40 ISSCR President Elaine Fuchs Closing Remarks 6:40 – 7:40 p.m. Closing Reception 800 Level – Plenary Hall Foyer Supported by Harvard Stem Cell Institute, Massachusetts General Hospital Center for Regenerative Medicine, and Stem Cell Program, Children’s Hospital Boston

35 Thank You to our 2011 Media Partners

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Exhibits and Exhibitor Floor Plan

37 ISSCR 9th Annual Meeting www.isscr.org

Exhibitor List

Company Name Booth Number Company Name Booth Number Abcam Plc 400 The Company of Biologists 404 Acris Antibodies 530 Corning Incorporated 700 Advanced Bioscience Resources 923 Custom Biogenic Systems 718 Advanced Microscopy Group 902 Cyntellect Inc 301 Agilent Technologies 838 Diamed Lab Supplies/Planer PLC 931 ALA Scientific Instruments 814 Dojindo Molecular Technologies Inc 824 AllCells LLC 325 DRVision Technologies LLC 1030 Alpha MED Scientific Inc 920 DV Biologics LLC 430 AlphaMed Press, Publisher of STEM CELLS® 619 DVS Sciences Inc 503 Amnis Corp 504 eBioscience 401 Applied Biological Materials Ltd 502 Elsevier 813 Applied StemCell Inc 428 EMD Millipore 919 Arteriocyte Inc 313 Epicentre Biotechnologies 305 ATCC 724 Esbe Scientific 1020 BD Biosciences 507 F1000’s The Scientist 803 Beckman Coulter Inc 909 Fisher BioServices 922 BioLamina AB 704 Fluidigm Corp 810 BioLegend 901 GE Healthcare 1008 Biological Industries Israel Beit Haemek Ltd 729 GeneTex Inc 819 BioMed Central Ltd 908 Genetica DNA Laboratories Inc 532 BioMedTech Laboratories Inc 632 GlobalStem 303 Bio-Rad Laboratories 825 Glycosan Biosystems Inc 533 BioSpherix Ltd 600 Guangzhou GZstem Biotechnologies Co Ltd 802 BioTime Inc 531 Hamilton Company 527 Blue Ocean Biomedical LLC 839 Hamilton Thorne Inc 410 BTX/Harvard Apparatus 833 HumanZyme Inc 929 Carl Zeiss MicroImaging 930 Huron Technologies 1002 CEDARLANE 1000 INDEC BioSystems 938 Cell Guidance Systems Ltd 500 Innovative Cell Technologies Inc 1022 Cell Line Genetics LLC 407 International Society for Cellular Therapy 720 Cell Press 811 KANEKA Corporation 403 Cell Signaling Technology 733 Kawasaki Heavy Industries Ltd 433 Cellartis AB 918 LABS Inc 603 CellGenix GmbH 1024 Leica Microsystems 630 Cellular Engineering Technologies Inc 323 Life Technologies 626 Celprogen Inc 311 Lonza 701,703 Center for IPS Cell Research and Application, Kyoto Univ 721 Mary Ann Liebert Inc 713 Chia Sheng Technology Co Ltd 1028 Massachusetts Human Stem Cell Bank & International Registry 722 Cold Spring Harbor Laboratory 912 Matrigen 1018

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Exhibitor List

Company Name Booth Number Company Name Booth Number McEwen Centre for Regenerative Medicine / RESBIO-NIH Funded Biomedical Technology Resource 629 University Health Network 801 Roche Diagnostics Corp 932 Miltenyi Biotec GmbH 418 SANYO 910 Molecular Devices Inc 800 Sarstedt Inc 605 MP Biomedicals 331 Scottish Development International (SDI) 823 MVE Chart 805 Sigma Life Science 903 National Center for Regenerative Medicine 739 Society for Neuroscience 821 National Institute of Biomedical Imaging and Bioengineering 711 Springer 728 Publishing Group 631 Stem Cell Network 625,623 The New York Stem Cell Foundation (NYSCF) 719 STEMCELL Technologies Inc 506 Nexcelom Bioscience 925 Stemgent 611 NOF America Corp 924 StemRD Inc 708 Norgen Biotek Corp 928 Sutter Instrument 408 Novus Biologicals 422 System Biosciences (SBI) 831 Olympus Canada Inc 431 TAP Biosystems 427 Ontario Stem Cell Initiative (OSCI) 710 Taylor Wharton 309 ORIGIO Inc 820 Thermo Scientific 830,832 Paragon Bioservices Inc 1012 Tissue Regeneration Therapeutics Inc 730 PeproTech Inc 526 TOKAI HIT Co Ltd 809 Phalanx Biotech Group 815 Tree Star Inc 812 Primorigen Biosciences Inc 705 Union Biometrica Inc 501 PromoCell GmbH 1004 VisualSonics Inc 723 Proteintech Group Inc 319 Vitrolife 702 QIAGEN Inc 709 WaferGen Biosystems 329 R&D Systems Inc 914 WiCell Research Institute 715 ReachBio LLC 601 Wiley-Blackwell 818 Regenerative Medicine, Future Medicine 804 Worthington Biochemical Corp 900 ReproCELL 829

39 40 www.isscr.org Final Program

Exhibitor/Supporter Directory

Abcam Plc - Booth 400 software and optical components for the life Alpha MED Scientific Inc - Dr. Seema Sharma sciences industry. The home of the EVOS® line Booth 920 of fully integrated all-in-one digital inverted 330 Cambridge Science Park Rika Yamazaki, Marketing Director microscopes, AMG is a concept-to-completion Cambridge, CB23 0WN company that provides innovative optical 209, 7-7-15, Saito-asagi United Kingdom solutions and products to research, clinical and Ibaraki, Osaka, 567-0085 Phone: +44-1223-696000 industrial applications all over the world. Japan [email protected] Phone: +81-72-648-7973 www.abcam.com Agilent Technologies – Booth 838 Fax: +81-72-648-7974 Abcam’s catalogue of more than 70,000 life Elaine Baccino, Life Sciences Campaign [email protected] science reagents includes cutting edge tools Coordinator www.med64.com for stem cells and developmental biology. We 2850 Centerville Road Our MED64 is a user-friendly micro-electrode have over 13,000 products in the stem cells and Wilmington, DE 19808 array system for in vitro electrophysiology. High- development field that include antibodies, marker USA quality extracellular signals are acquired with the panels, active proteins, lysates and ELISA’s. industry’s lowest-noise system. It is a powerful Visit www.abcam.com/stemcells. Phone: +1-302-636-3558 [email protected] solution for drug screening with stem cell-derived cardiomyocytes and neurons. Acris Antibodies - Booth 530 www.agilent.com/chem/genomics John Mountzouris, President Agilent Technologies, a leading supplier of life AlphaMed Press, Publisher of science instrumentation and reagents, with 6815 Flanders Drive STEM CELLS® - Booth 619 San Diego, CA 92121 solutions optimizing workflow productivity and performance through automation. From sample Sharon Lee, Marketing and USA preparation to data analysis, Agilent provides Communications Manager Phone: +1-858-888-7900 microarrays, reagents, real-time PCR and Next 318 Blackwell St, Suite 260 Fax: +1-858-888-7904 Gen Sequencing. Durham, NC 27701 [email protected] USA www.acris-antibodies.com ALA Scientific Instruments - Phone: +1-919-680-0011 Antibodies for stem cell research, neuroscience, Booth 814 Fax: +1-919-680-4411 cancer, development and immunology for life Margaret Badon, Office Manager [email protected] science researchers. 60 Marine St http://journals.alphamedpress.com/ Farmingdale, NY 11735 AlphaMed Press, publisher of: STEM CELLS Advanced Bioscience Resources - USA and STEM CELLS TRANSLATIONAL MEDICINE Booth 923 Phone: +1-631-393-6401 (SCTM). STEM CELLS focuses on stem and Perrin Larton, Procurement Manager Fax: +1-631-393-6407 progenitor cell biology. SCTM bridges research Procurement Dept [email protected] and clinical trials, focusing on clinical utilization of 1516 Oak St #303 www.alascience.com stem cell molecular and cellular biology. Alameda, CA 94501 ALA Scientific and MultiChannel Systems Amnis Corporation - Booth 504 USA develop recording and perfusion instruments for Phone: +1-510-865-5872 electrophysiology. Leading stem cell researchers Bob Smith-McCollum, Marketing Specialist Fax: +1-510-865-4090 use our Mulitelectrode Array (MEA) system 2505 Third Ave, Suite 210 [email protected] to electrophysiologically characterize stem Seattle, WA 98121 www.abr-inc.com cell derived cardiomyocytes and neurons. The USA MEA System is easy to use and reveals reliable Phone: +1-206-374-7000 Advanced Bioscience Resources procures and electrophysiological information on stem cell Fax: +1-206-576-6895 distributes human fetal Tissue, full term umbilical derived cardiomyocytes and neurons. cord blood, and adult peripheral blood for [email protected] research. ABR can also provide tissue under www.amnis.com AllCells LLC - Booth 325 cGTP guidelines. Amnis introduces the revolutionary new John Chang, Marketing FlowSight, a 12 channel flow cytometer that Advanced Microscopy Group - 5858 Horton St #360 is priced for every lab. For more demanding Booth 902 Emeryville, CA 94608 requirements, Amnis’ ImageStream imaging flow USA Niki Amato, Marketing & International Sales cytometer delivers the ultimate in image quality Phone: +1-510-450-3900 and image-based applications. 22025 20th Ave SE, Suite 100 [email protected] Bothell, WA 98034 www.allcells.com USA Phone: +1-425-368-0444 AllCells serves as a reliable provider of more Fax: +1-425-368-0555 than 150 primary cell types and related RNA & [email protected] cDNA from normal and disease-state tissues. Cell types include stem cells (progenitor, CD34+, www.amgmicro.com etc.), hematopoietic, human hematologic disease, Focused on quality, performance and innovation, endothelial, dendritic, HUVEC cells and many AMG (Advanced Microscopy Group) designs and others. develops state-of-the-art microscopy equipment, 41 ISSCR 9th Annual Meeting www.isscr.org

Exhibitor/Supporter Directory

Applied Biological Materials Ltd - ATCC - Booth 724 Beckman Coulter Inc - Booth 909 Booth 502 Robin Rothrock, Market Development Carol Huggins, Sr. Exhibit Specialist Peter Li, Chief Scientific Officer Director 250 S Kraemer Blvd 8-13520 Crestwood Place 10801 University Blvd Brea, CA 92822 Richmond, BC V6V 2G2 Manassas, VA 20110 USA Canada USA Phone: +1-714-993-5321 Phone: +1-778-588-0818 Phone: +1-703-365-2700 www.beckmancoulter.com Fax: +1-604-247-2414 Fax: +1-703-365-2750 Beckman Coulter delivers comprehensive system [email protected] [email protected] solutions that address the multiple steps needed www.abmgood.com www.atcc.org to process stem cell development. We collaborate with you to understand your requirements and ABM is a premier, one-stop supplier for ATCC introduces a new iPS cell repository, create flexible solutions that meet your evolving innovative genetic and cellular reagents for established to accession and globally distribute needs. We offer a broad technology portfolio and the life sciences and biotech community. Our highly qualified, standardized cell lines various bundling options that will help your lab affordable products and services include 20,000 reprogrammed via expression of OSKM - without deliver results accurately and efficiently. reliable antibodies, miRNA profiling, and high gene integration. A novel media formulation performance qPCR, PCR, transfection and viral simplifies pluripotent cell research. expression reagents. BioLamina AB - Booth 704 Australian Therése Kallur, Kristian Tryggavson, Yi Sun Applied StemCell Inc - Booth 428 Stem Cell Centre Bronze Löfströms Allé 5 Sundbyberg, 172 66 Makiko Ogawa, Marketing Manager Rebecca Skinner, Supporter Sweden 1165 O’Brien Drive, Suite A Senior Manager – Phone: +46-8-5888-5180 Menlo Park, CA 94025 Communications and Networking Fax: +46-8-5198-9288 USA Ground Floor, Building 75 [email protected], kristian@ Phone: +1-408-773-8007 Monash University biolamina.com, [email protected] Fax: +1-408-773-8238 Wellington Road [email protected] Clayton, Melbourne VIC 3800 BioLamina is manufacturing and selling purified www.appliedstemcell.com Australia human recombinant laminin isoforms that are Phone: +61-3-9271-1100 completely defined and animal protein-free, to be Applied StemCell, Inc. has unique solutions for used as substrate solutions for cell cultures all the Fax: +61-3-9271-1199 your stem cell research. Please contact us to way from bench to bedside. request updated information for customized [email protected] site-specific transgenic services, in vivo teratoma www.stemcellcentre.edu.au analysis and more. Visit www.appliedstemcell. BioLegend - Booth 901 com to learn about our custom services/research The ASCC was founded in 2002 to take 11080 Roselle St products. advantage of Australia’s significant strengths in San Diego, CA 92121 the field of stem cell research. It funds groups USA across Australia investigating all types of stem Arteriocyte Inc - Booth 313 cells and has a strong focus on public education, Phone: +1-858-455-9588 Ray Sodha, Project Scientist partnering and networking. [email protected] 7100 Euclid Ave, Suite 150 www.biolegend.com Cleveland, OH 44103 BD Biosciences - Booth 507 Highest quality fluorochrome conjugated USA Julia Lizondo, Product Manager antibodies for flow cytometry and Phone: +1-216-456-9646 10975 Torreyana Road immunofluorescence. Special discounted pricing Fax: +1-216-456-9641 San Diego, CA 92121 for bulk orders on biofunctional LEAF™ (Low- Platinum Endotoxin, Azide-Free) antibodies. Cutting-edge [email protected] USA www.nanex.com Supporter LEGEND MAX™ ELISA Kits with pre-coated Phone: +1-877-232-8995 plates. Comprehensive Chemokine Receptor Ab Arteriocyte Inc. is an independent company Fax: +1-858-812-3739 Panel. Functional recombinant cytokines. focused on developing and commercializing [email protected] cell-based products and medical devices. www.bdbiosciences.com/stemcells Biological Industries Israel Beit The company has a developing portfolio of technologies addressing unmet research and BD Biosciences is a world leader in bringing Haemek Ltd - Booth 729 clinical needs, including NANEX and Magellan innovative clinical diagnostic and research tools Gilad Landes, VP Marketing & Sales product families. to life scientists, clinical researchers, laboratory Kibbutz Beit Haemek, 25115 professionals and clinicians who are involved in Israel basic research, drug discovery and development, biopharmaceutical production and disease Phone: +972-4-9960594 management. Fax: +972-4-9968896 [email protected]; [email protected] www.bioind.com BI- Biological Industries manufactures and supplies to biopharmaceutical companies, research and cytogenetic laboratories worldwide products for cell culture and . 42 www.isscr.org Final Program

Exhibitor/Supporter Directory

Our stem cell portfolio STEMsySTEM™ includes BioSpherix Ltd - Booth 600 BTX/Harvard Apparatus - unique products for hESC, iPSC, MSC culture and Kevin Murray, Director Of Sales Booth 833 other novel animal component-free products. 19 Demott St Derek Palmer, Sales Manager Lacona, NY 13083 84 October Hill Road BioMed Central - Booth 908 USA Holliston, MA 01746 Nishkala Thiru, Marketing Executive Phone: +1-315-387-3414 USA 236 Grays Inn Road Fax: +1-315-387-3415 Phone: +1-800-272-2775 London, WC1X 8HB [email protected] Fax: +1-508-429-5732 United Kingdom www.biospherix.com [email protected] Phone: +44-203-192-2009 www.btxonline.com Fax: +44-203-192-2010 BioSpherix exhibits the world’s first and only point-of-care cGMP compliant stem cell [email protected] BTX is introducing the new BTXpress incubation and processing systems. Any cell www.biomedcentral.com Electroporation Solution, to increase transfection processing application: explants or implants; efficiency and viability of cell lines and difficult BioMed Central publishes more than 200 peer- minimal or maximal manipulation; autologous or to transfection cells. BTX provides the tools and reviewed open access journals, including Stem allogeneic cells; pre or post biorepository. Learn technologies needed to transfect mammalian Cell Research & Therapy. Find out more about more at booth #600. cells, animal tissues and fusion of cells. BioMed Central and open access publishing, and collect your copy of Stem Cell Research & BioTime, Inc - Booth 531 Carl Zeiss MicroImaging LLC - 930 Therapy at booth 908. Barbara Domingo, Director, Marketing & Karin Salerno, Marketing Communications Communications Manager BioMedTech Laboratories Inc - 1301 Harbor Bay Parkway One Zeiss Drive Booth 632 Alameda, CA 94502 Thornwood, NY 10594 Joachim Sasse, President USA USA 3802 Spectrum Blvd, Suite 154 Phone: +1-510-521-3390 Phone: +1-800-233-2343 Tampa, FL 33612 [email protected] Fax: +1-914-681-7446 USA www.biotimemail.com [email protected] Phone: +1-813-985-7180 BioTime is a biotechnology company that www.zeiss.com/micro Fax: +1-813-558-2000 develops and markets products in the field of Carl Zeiss sells and supports a broad range of [email protected] stem cells and regenerative medicine. research microscopes, laser microdissection www.biomedtech.com systems, digital cameras and image analysis Culture surfaces optimized for specific cell types, Blue Ocean Biomedical LLC – software. including stem-cells and neural-cells are featured. Booth 839 BioMedTech high-quality coatings (PDL, PLL, Willy Gutierrez, Director of Marketing CEDARLANE - Booth 1000 Collagen, Gelatin, BME, Laminin, Fibronectin, 1 SW 129th Ave, Suite 201 Naomi Wessel-Bonnar, Associate Marketing Laminin/Collagen IV) are expertly applied to all Pembroke Pines, FL 33027 Manager imaging plates, microplates, flasks, dishes, bio- 4410 Paletta Court production vessels and roller-bottles. USA Phone: +1-954-404-6002 Burlington, ON L7L 5R2 Canada Bio-Rad Laboratories - Booth 825 Fax: +1-954-432-7161 Phone: +1-800-268-5058 Marcie Wright, Marketing Manager [email protected] Fax: +1-289-288-0020 2000 Alfred Nobel Drive www.blueoceanbio.com [email protected] Hercules, CA 94547 Blue Ocean Biomedical specializes in the www.cedarlanelabs.com USA development of innovative, automated, load & Phone: +1-510-741-1000 go™ cell analysis systems for immune monitoring CEDARLANE® (ISO 13485 registered) specializes Fax: +1-510-741-5630 and a variety of flow cytometry applications. Blue in providing high quality research & diagnostic reagents to the life science community. We offer [email protected] Ocean systems are designed for moderate-to- high demand applications where tests results are one of the most extensive product listings both http://discover.bio-rad.com desired in less time, with greater efficiency and at domestically & internationally. Delivering Today’s Bio-Rad Laboratories has played a leading role lower costs. Innovations for the Science of Tomorrow™ in the advancement of scientific discovery for more than 50 years by providing a broad range of innovative products and services to the life science research and clinical diagnostic markets. Founded in 1952, Bio-Rad serves research and industry customers around the world through its global network of operations.

43 ISSCR 9th Annual Meeting www.isscr.org

Exhibitor/Supporter Directory

Cell Guidance Systems - Cell Signaling Technology - Cellular Engineering Technologies Booth 500 Booth 733 Inc – Booth 323 Michael Jones, CEO Carrie Ann Brown, Trade Show Manager Anant M. Kamath, Chief Operating Officer 2 Munro House 3 Trask Lane 2500 Crosspark Road, Suite E232 Trafalgar Way Danvers, MA 01923 Coralville, IA 52241 Bar Hill, Cambridge, CB23 8SQ USA USA United Kingdom Phone: +1-978-867-2300 Phone: +1-319-665-3000 Phone: +44-1954-785-007 Fax: +1-978-867-2400 Fax: +1-319-665-3003 [email protected] [email protected] [email protected] www.cellgs.com www.cellsignal.com www.celleng-tech.com Cell Guidance Systems develops and sells Cell Signaling Technology develops the highest Cellular Engineering Technologies (CET) is a reagents including: quality antibodies, cytokines and related reagents leading provider of adult somatic stem cells, 1. Pluripro® synthetic media for maintenance for the study of stem cell signaling, including our human and murine IPS cells, primary cells, growth of human pluripotent cells exclusive line of XP™ monoclonal antibodies. and differentiation media. CET provides cells from 2. Standard growth factors with high biological Please visit our website at www.cellsignal.com to normal volunteers and donors who have specific activity view our entire product line. clinical conditions. 3. Coming soon: STAR growth factors with *unprecedented* levels of biological activity Cellartis AB - Booth 918 Celprogen Inc - Booth 311 Dr. Barry Middleton, Head of Business Jay Sharma, CEO/CSO Cell Line Genetics - Booth 407 Development 1871 N Gaffey St, Suite A & B Robert Margolin, Director of Marketing 1 Würzburg Court San Pedro, CA 90731 510 Charmany Dr, Suite 254 Dundee, DD2 1FB USA Madison, WI 53719 United Kingdom Phone: +1-310-547-3975 USA Phone: +44-1382-56-99-70 Fax: +1-310-547-2975 Phone: +1-646-415-9265 Fax: +44-1382-56-82-42 www.celprogen.com Fax: +1-212-591-6025 [email protected] [email protected] Celprogen is a biotechnology company that www.cellartis.com focuses on stem cell research and therapeutics. www.clgenetics.com Cellartis has 10 years of experience of stem Celprogen has three divisions; life sciences, Cell Line Genetics is recognized as the leader cell research and development focused on therapeutics and devices. The life science division in stem characterization, identification and cell hepatocytes and cardiomyocytes for drug provides stem cell research tools including stem line quality-assurance products and services. discovery and also as a partner in regenerative cells, progenitor cells, iPCs, media (differentiation, The company’s scientific team has 50+ years of medicine. We offer stem cell derived specialised expansion & un-differentiation) ECM, cell based combined experience and has tested more than cells and can also work in collaboration with assays and cancer stem cells. For therapeutics we 6,000 stem cell lines from 10 species. industrial partners. have several drugs that are in their pre-clinical phase for the treatment of cardiovascular disease, Cell Press – Booth 811 CellGenix GmbH - Booth 1024 diabetes, neurology and liver disease. Also for services we provide various drug screening assays Jonathan Atkinson, Director of Marketing Richard Neubiser, GM/VP CellGenix US for drug discovery programs inculding in vivo and 600 Technology Square, 5th Floor Operations in vitro systems for pharmaceutical companies. Cambridge, MA 02139 303 Main St, 100-C USA Antioch, IL 60002 Celsense Inc Phone: +1-617-661-7057 Silver USA Charles F. O’Hanlon, III, President and CEO Fax: +1-617-661-7061 Phone: +1-847-395-7277 603 Stanwix St., Suite 348 [email protected] Supporter Fax: +1-847-395-0808 Pittsburgh, PA 15222 www.cell.com [email protected] USA Bronze Visit Cell Press during ISSCR and pick up free www.cellgenix.com Phone: +1-412-263-2870 Supporter samples of leading high impact journals including CellGenix GmbH is an innovative Fax: +1-412-263-2878 Cell (stem cells special issue); Cell Stem Cell (June biopharmaceutical company headquartered [email protected] issue with focus on regenerative medicine) and in Freiburg, Germany. CellGenix develops, www.celsense.com Neuron (neural stem cells special issue), and take manufactures and markets cGMP cell and protein the opportunity to meet the Editors of Cell Stem therapeutics for cancer and orthopedic patients, Celsense, Inc. offers products used to non- Cell and Cell. as well as high-quality reagents for therapeutic ex invasively visualize and measure the trafficking of vivo cell processing. cells in vivo using magnetic resonance imaging. Applications include the development of therapeutic agents, and monitoring the delivery of therapeutic cells in clinical trials and approved use.

44 www.isscr.org Final Program

Exhibitor/Supporter Directory

Centre for Commercialization of Children’s Hospital Boston Corning Incorporated - Booth 700 Regenerative Medicine Leonard I. Zon, Director, Stem Cell Program Dawn Jackson, Product Line Manager Michael H. May, PhD, Chief Executive 300 Longwood Ave. Advanced Cells and Surfaces Officer Boston, MA 02115 900 Chelmsford St 160 College St, Suite 1110 USA Silver Tower 2, 4th Floor Toronto, ON M5S 3E1 Phone: +1-617-919-2069 Supporter Lowell, MA 01851 Canada Bronze Fax: +1-617-730-0222 USA Phone: +1-416-978-3751 Supporter [email protected] Phone: +1-800-492-1110 or [email protected] http://stemcell.childrenshospital.org/ +1-978-442-2200 www.ccrm.ca The mission of the Stem Cell Program at Fax: +1-978-442-2476 [email protected] The Centre for Commercialization of Regenerative Children’s Hospital Boston is to explore and Medicine (CCRM) is a Canadian not-for-profit understand the promise of stem cell biology as www.corning.com/lifesciences dedicated to supporting the development of a key to treatments for disease, and to translate Corning Life Sciences, a global leader in foundational technologies that accelerate the that promise into effective clinical therapies. the laboratory products market, offers commercialization of stem cell and biomaterials- core capabilities in materials, surfaces and based technologies and therapies. CCRM is Cold Spring Harbor Laboratory - biophotonics. Products include new surfaces such a public, private collaboration representing Booth 912 as Synthemax™ Surface for stem cell culture, six world class academic institutions and a 16 Mala Mazzullo, Executive Assistant to the Ultra Low Attachment Surface, Transwell® company industry consortium. Publisher Permeable Supports and our CellBIND® Surface. One Bungtown Road Center for IPS Cell Research and Cold Spring Harbor, NY 11724 Custom Biogenic Systems - Application (CiRA), USA Booth 718 Kyoto Univ - Booth 721 Phone: +1-516-422-4005 Cass Schumann, Sales/Marketing Director Akemi Nakamura, International Public Fax: +1-516-422-4092 150 Shafer Drive Communications Office [email protected] Romeo, MI 48065 53 Kawahara-Cho www.cshl.edu and www.cshlpress.com USA Shogoin, Sakyo-Ku Phone: +1-800-523-0072 Cold Spring Harbor Laboratory continues to Fax: +1-586-331-2588 Kyoto, 606-8507 shape contemporary biomedical research and Japan education with programs in cancer, neuroscience, [email protected] Phone: +81-75-366-7005 plant biology and quantitative biology. Its www.custombiogenics.com Fax: +81-75-366-7024 Meetings & Courses program hosts more than Displayed will be a complete range of equipment [email protected] 8,000 scientists from around the world each for bone marrow, cord blood and stem cell year and its Press publishes books, journals and CiRA was established in 2008 at Kyoto University cryopreservation. The - 190 C ISOTHERMAL electronic media for scientists, students and the sample storage freezers, controlled rate freezing in Kyoto, Japan, as the world’s first institute that general public. focuses on iPS cell research. Dr. Shinya Yamanaka, systems, and Ln2 dry shipping containers. Also the pioneer of iPS cell technology, serves as the displayed will be canister and frame inventory CiRA director. The Company of Biologists - systems for storing bags along with rack and box Booth 404 systems for storing tubes. Chia Sheng Technology Co., Ltd - Sarah Sharpe, Marketing Associate Booth 1028 140 Cowley Road Peter Chiu, Sales Manager Bidder Bldg 4F-1 No 262 Sec 2 Henan Road Cambridge, CB4 0DL Xitun District United Kingdom Taichung City, 407 Phone: +44-1223-433319 Taiwan R.O.C. Fax: +44-1223-423353 Phone: +886-4-627071236 [email protected] Fax: +886-4-27071246 www.biologists.com [email protected] The Company of Biologists is the not-for-profit www.cst.org.tw publisher of the well established, international journals Development, Disease Models & CS Technology is an equipment company Mechanisms, Journal of Cell Science and The specialized in manufacturing precision instruments Journal of Experimental Biology. for industrial and scientific applications.

45 ISSCR 9th Annual Meeting www.isscr.org

Exhibitor/Supporter Directory

Cyntellect, Inc - Booth 301 Dojindo Molecular Technologies, DVS Sciences Inc - Booth 503 Caitlyn Borst, Marketing and Inc - Booth 824 Scott Tanner, President & CEO Communications Specialist Taro Nagata, Sales & Technical Support 12-70 Esna Park Drive 6620 Mesa Ridge Road 30 West Gude Drive, Suite 260 Markham, ON L3R 6E7 San Diego, CA 92121 Rockville, MD 20850 Canada USA USA Phone: +1-905-513-1704 Phone: +1-858-875-1600 Phone: +1-301-987-2667 [email protected] Fax: +1-858-875-1610 Fax: +1-301-987-2687 www.dvssciences.com [email protected] [email protected] DVS Sciences Inc. is an analytical equipment and www.cyntellect.com www.dojindo.com reagents development company that produces Cyntellect creates and commercializes best-in- Cell based assay, cell proliferation / cytotoxicity the CyTOF™ mass cytometer for individual cell class products to analyze and process living cells assay, transfection reagent, oxidative stress analysis based on elemental mass-spectrometry more efficiently and with far greater accuracy detection, protein detection, labeling kits for IgG detection technology, and the MAXPAR™ system than any technology in use today. Cyntellect’s and protein, DNA/RNA isolation kit. of novel reagents related to massively multi- products are currently used to advance the parametric biological assays. state of life science research, biopharmaceutical DRVision Technologies LLC - production, stem cell research and drug eBioscience - Booth 401 discovery. The Company’s in situ microplate- Booth 1030 Edwin Chau, Associate Director based cytometry systems process cells with great 15921 NE 8th St., Suite 200 10255 Science Center Drive precision and simplicity in their natural state, Bellevue, WA 98008 San Diego, CA 92121 allowing cells to maintain their proper physiologic USA USA behavior. Cyntellect’s expanding cellular analysis Phone: +1-425-653-5589 Phone: +1-888-999-1371 and processing portfolio is expected to play an Fax: +1-425-746-0859 enabling role in the coming age of advanced cell- Fax: +1-858-642-2046 [email protected] based diagnostics and therapeutics. For additional [email protected] www.svcell.com information please visit www.cyntellect.com. eBioscience is committed to developing and DRVision is proud to launch the Stem Cell manufacturing high-quality, innovative reagents Development Analyzer suite for common stem cell applications in an ISO certified facility. As a provider of Sarah Sharpe, Marketing Associate in quantitative microscopy. Consisting of SVCell more than 10,000 products, we empower our software and application recipes, it provides Bidder Bldg., 140 Cowley Road customers worldwide to obtain exceptional results exceptional fluorescence and brightfield image Cambridge CB4 0DL by using reagents that offer a new standard of pattern analysis capability in a stand-alone excellence in the areas of innovation, quality and United Kingdom Bronze software package. value. Phone: +44-1223-433319 Supporter Fax: +44-1223-423353 DV Biologics LLC – Booth 430 Elsevier Inc – Booth 813 [email protected] Janna Lacher, Business Development Andrea Cowan, Marketing Manager dev.biologists.org Manager 525 B Street, Suite 1800 Development is a leading primary research journal 1239 Victoria St. San Diego, CA 92101 in the field of developmental biology that is Costa Mesa, CA 92627 USA published by The Company of Biologists, a not- USA Phone: +1-619-699-6539 for-profit charitable organization run by biologists Phone: +1-888-773-5959 Fax: +1-619-699-6380 for the benefit of the biological community. Visit Fax: +1-877-773-5959 Development’s homepage dev.biologists.org [email protected] [email protected] www.elsevierdirect.com/stemcells www.dvbiologics.com Diamed Lab Supplies Inc – Elsevier is pleased to publish Stem Cells, Scientific Booth 931 DV Biologics is a global supplier of human Facts and Fiction, by Mummery, Wilmut, Van biological tools including normal and disease- De Stolpe and Roelen. Using straightforward Derek Johnson, President specific cells, stem cells, media, proteomic language it explains the basic biology of stem 3069 Universal Drive tools and custom cell products. DV Biologics cells and their applications in modern medicine Mississauga, ON L4X 2E2 manufactures only the highest quality products and future therapy. Canada for scientists engaging in cell based studies. Phone: +1-800-434-2633 Fax: +1-905-625-6280 [email protected] www.diamed.ca Large selection of lab consumables, examination gloves, equipment. service and maintenance programs available.

46 www.isscr.org Final Program

Exhibitor/Supporter Directory

EMD Millipore - Booth 919 F. Hoffman-La Roche Ltd Fisher BioServices - Booth 922 Natasha Sotomayor, Exhibits Specialist Dr. Matthias Steger, Global Head, Research Megan Collins, Marketing and Sales 290 Concord Road & Technology Partnering Coordinator Billerica, MA 01821 Grenzacherstrasse 124 14665 Rothgeb Drive USA Bronze Basel 4070 Bronze Rockville, MD 20850 Phone: +1-978-715-1483 Supporter Switzerland Supporter USA Fax: +1-978-715-1393 Phone: +41-61-687-9544 Phone: +1-301-315-8460 [email protected] Fax: +41-61-688-7990 Fax: +1-301-838-9320 www.emd-millipore.com [email protected] [email protected] EMD Millipore is the life science division of www.roche.com www.fisherbioservices.com Merck KGaA, Germany, supporting research, Roche is a leader in research-focused healthcare Fisher BioServices, a business unit of Thermo development and production of biotech and with strengths in pharmaceuticals and diagnostics. Fisher Scientific, is a professional cell therapy pharmaceutical therapies. Our complete solutions Focusing in oncology, virology, inflammation, services business. With locations around for advanced stem cell analysis feature Scepter™, metabolism and CNS. Roche also leads in in vitro the world, we are committed to providing the first handheld, automated cell counter and diagnostics, tissue-based cancer diagnostics and is biorepository, laboratory, and clinical trial services guava® integrated benchtop flow cytometry a pioneer in diabetes management: and logistics infrastructure to the stem cell and systems. www.roche.com. cell therapy industries. Our global facilities and capabilities provide integrated solutions EPICENTRE Biotechnologies - Fate Therapeutics Inc. from collection site or CMO to repository and clinical site. Services include GTP biorepository, Booth 305 Scott Wolchko, Chief Financial Officer biobanking, laboratory services, cell or specimen Fraz Syed, Product Manager 3535 General Atomics Court, Suite 200 collection kits, on-site inventory management and 726 Post Road San Diego, CA 92121 cryogenic and ultra low cold chain management. Madison, WI 53713 USA General USA Phone: +1-858-875-1800 Fluidigm Corporation - Booth 810 Phone: +1-800-284-8474 Supporter Fax: +1-858-875-1843 7000 Shoreline Court, Suite 100 [email protected] [email protected] South San Francisco, CA 94080 EPICENTRE features innovative kits for preparing www.fatetherapeutics.com USA mRNA-Seq, miRNA-Seq and Next-Gen genomic Fate Therapeutics is interrogating stem cell Phone: +1-650-266-6000 DNA sequencing libraries. Kits enabling hundreds biology to develop therapeutics based on Fax: +1-650-871-7152 of sensitive qRT-PCR reactions and transcription modulating cell fate and to enable a new drug [email protected] profiling microarray studies from as little as one discovery paradigm which includes proprietary iPS www.fluidigm.com cell will also be highlighted. cell technology. The company’s first therapeutic candidate is in Ph1b for hematopoietic Fluidigm develops, manufactures and markets ESBE Scientific - Booth 1020 reconstitution. life-science systems based on integrated fluidic Bob Dubasz, Director of Sales and circuits (IFCs). This technology furthers research by minimizing costs and enhancing sensitivity for Marketing F1000’s The Scientist - Booth 803 applications such as single-cell gene expression 80 McPherson St Stephanie Eaves, Marketing Coordinator profiling, high-throughput SNP genotyping, and Markham, ON L3R 3V6 121 West 27th St., Suite 604 next-generation sequencing. Fluidigm products Canada New York, NY 10001 are used for research only. Phone: +1-905-475-8232/+1-800-268-3477 USA Fax: +1-905-475-5688/+1-800-387-0476 Phone: +1-800-258-6008 GE Healthcare - Booth 1008 [email protected] [email protected] 800 Centennial Ave www.esbe.com www.the-scientist.com PO Box 1327 ESBE Scientific, a completely Canadian laboratory The Scientist, F1000’s magazine of the life Piscataway, NJ 08855-1327 supply company is exhibiting products for the sciences, provides compelling print and online Phone: +1-800-526-3593 or +1-732-457- biotechnology and medical laboratories. Products coverage of the latest developments in research, 8000 on display include BAKER biosafety cabinets, technology, careers and business. Fax: +1-877-295-8102 RUSKINN In Vivo Workstations, NEW SANYO [email protected] technology ultra-low freezers, incubators as well www.gelifesciences.com as general laboratory apparatus. GE Healthcare Life Sciences provides tools for drug discovery and cellular technologies so research scientists and specialists worldwide can be more productive, effective and creative. Our vision is to be the partner of choice in cell and protein research and the leader in life sciences services.

47 ISSCR 9th Annual Meeting www.isscr.org

Exhibitor/Supporter Directory

GeneTex Inc - Booth 819 GlobalStem - Booth 303 Hamilton Company - Booth 527 Fawn Sammy Andre Lubarsky, Director of Sales Kelli Cavallaro, Sales Administration 2456 Alton Parkway 9430 Key West Ave, Suite 130 Manager Irvine, CA 92606 Rockville, MD 20850 4970 Energy Way USA USA Reno, NV 89502 Phone: +1-877-GENETEX Phone: +1-301-545-0238 USA Fax: +1-949-309-2888 Fax: +1-301-424-1989 Phone: +1-775-858-3000 [email protected] [email protected] Fax: +1-775-858-3024 www.genetex.com www.globalstem.com [email protected] GeneTex is a high quality antibody manufacturer GlobalStem provides high-quality, standardized www.hamiltonrobotics.com and supplier with more than 30,000 antibodies reagents and contract services to support stem Hamilton Robotics is dedicated to the design covering various research disciplines. We offer cell and CNS research. Our products include and manufacture of automated liquid handling the highest quality antibody reagents following stem cell culture reagents, stem cells and neural workstations. Key to our products is our extensive research, development and multiple cell types derived from pluripotent stem cells. air displacement pipetting and monitoring species/application validation. We provide contract services for expanding, technology and software. Our workstations serve banking and characterizing stem cells to ensure as a high precision base upon which to provide Genetica DNA Laboratories, Inc - consistency and reliability for research and drug automated solutions. Booth 532 discovery. Alexander Sinelnikov, PhD, Assistant Lab Hamilton Thorne, Inc - Booth 410 Director Glycosan BioSystems, Inc - Lily Reed, Director of Research Market and 8740 Montgomery Road Booth 533 Applications Cincinnati, OH 45236 Thomas Zarembinski, Director 100 Cummings Center, Suite 465E USA 675 Arapeen Drive, Suite 302 Beverly, MA 01915-6143 Phone: +1-513-985-9777 Salt Lake City, UT 84108 USA Fax: +1-513-985-9983 USA Phone: +1-978-921-2050 [email protected] Phone: +1-801-583-8212 Fax: +1-978-921-0250 www.celllineauthentication.com Fax: +1-801-588-0497 [email protected] [email protected] www.hamiltonthorne.com Genetica DNA Laboratories, Inc. has been www.glycosan.com providing DNA testing services in the areas of Hamilton Thorne presents its laser systems human DNA identity and DNA family relationship Glycosan BioSystems sells synthetic and natural for stem cell derivation and propagation. The testing for more than 20 years. We now offer hydrogels for stem cell culture and implantation. XYClone® features multi-pulse Staccato® reliable, fast, and affordable authentication software for ICM excision or biopsy for PGD. of human cancer cell lines, stem cell lines and Guangzhou GZstem The Stiletto™ “light-knife” offers rapid isolation, autoscoring and/or ablation of cultured cells xenografts to research facilities within the United Biotechnologies Co., Ltd - States and throughout the world. during passaging. Booth 802 Genome Canada Violla (Man) Dai, Operating Manager Harvard Stem Cell Institute Hélène Meilleur, Director of D1016 Int’l Business Incubator, Science City Maureen Herrmann, Program and Communications Guangzhou, 510663 Administrative Manager 150 Metcalfe St, Suite 2100 China Holyoke Center, Suite 727W Ottowa, ON K2P 1P1 Phone: +86-20-32299967 or 1350 Massachusetts Ave. General +86-18688887717 Canada Platinum Cambridge, MA 02138 Silver Phone: +1-613-751-4460 Supporter Fax: +86-20-32299957 USA Supporter Fax: +1-613-751-4474 [email protected] Supporter Phone: +1-617-496-6647 [email protected] www.gzstem.com.cn Fax: +1-617-496-6625 www.genomecanada.ca The mission of the company is the development [email protected] of human induced pluripotent stem cells (iPSCs), www.hsci.harvard.edu Genome Canada is a not-for-profit corporation derived from primary cell or tissue biopsy of dedicated to developing and implementing a HSCI is a scientific collaborative aimed at fulfilling normal and diseased human specimens. Also, the national strategy in genomics and proteomics the promise of stem cells. expansion of optimized protocols for directed research for the benefit of all Canadians. differentiation of normal and diseased human iPSCs into different cell lineages.

48 www.isscr.org Final Program

Exhibitor/Supporter Directory

The Hospital for Sick Children, INDEC BioSystems - Booth 938 Johnson & Johnson Research Institute 4701 Patrick Henry Drive, Building 24 Garry Neil, Corporate VP, Science & Janet Rossant, Chief of Research Santa Clara, CA 95054 Technology 555 University Ave. USA 410 George St. Toronto, ON M5G 1X8 Phone: +1-408-986-1600 New Brunswick, NJ 08901 Canada Bronze Fax: +1-408-986-1605 USA Gold Phone: +1-416-813-6577 Supporter [email protected] Phone: +1-732-524-1173 Supporter Fax: +1-416-813-5085 www.indecbiosystems.com Fax: +1-732-524-2549 [email protected] Modular solutions for fluorescence imaging – in [email protected] www.sickkids.ca/research vivo; in vitro. www.jnj.com SickKids Research Institute is one of Canada’s In-chamber magnification, real-time spectral Johnson & Johnson is the world’s most leading hospital-based research institutes. We separation. comprehensive and broadly based manufacturer undertake research from molecules to stem cells FluorVivo 100/300 – real-time, color, multi-label, of health care products, as well as a provider of to disease models, with the goal of improving CFP/GFP to near-IR. related services, for the consumer, pharmaceutical children’s health worldwide. Our vision: Healthier FluorVivo 1000 – monochrome imaging. and medical devices and diagnostics markets. Children. A better world. FluorVivo nIR – infrared imaging. The more than 250 Johnson & Johnson operating FluorVivo Mag – in vivo microscopy. companies employ approximately 115,500 people FluorVivo Surgery – preclinical fluorescence- in 60 countries throughout the world. HumanZyme, Inc - Booth 929 guided surgery. Fredric Rhoads, VP of Sales and Marketing Juvenile Diabetes Research 2201 W Campbell Park Drive, Suite 24 Innovative Cell Technologies Inc - Foundation (JDRF) Chicago, IL 60612 Booth 1022 USA Adrianne Wong, Senior Scientific Program C. Kevin Becker Phone: +1-312-738-0127 Manager - Beta Cell 6790 Top Gun St, #1 Fax: +1-312-738-0136 26 Broadway, 14th Floor San Diego, CA 92121 General [email protected] New York, NY 10004 USA www.humanzyme.com USA Supporter Phone: +1-858-587-1716 Phone: +1-212-479-7674 HumanZyme, Inc. provides a range of superior Fax: +1-858-453-2117 Fax: +1-212-785-9609 human protein cytokines, phosphatases and [email protected] [email protected] coagulation proteins that conventional production www.innovativecelltech.com methods can not produce. HumanZyme uses its www.jdrf.org proprietary engineered human cells, expression ICT manufactures and sells Accutase, Cell JDRF is a leader in setting the agenda for diabetes vectors, and growth medium to ensure high-yield Detachment Solution and Accumax, Cell research worldwide, and is the largest charitable production of authentic recombinant human Dissociation Solution. These products are funder and advocate for type 1 diabetes research. proteins with natural human post-translational superior replacements for trypsin and collagenase. The mission of JDRF is to find a cure for diabetes modifications, such as phosphorylation and Stop by our booth for a free sample. and its complications. glycosylation. The company also provides timely and cost-effective contract production of International Society for Cellular KANEKA Corporation - Booth 403 recombinant human proteins from human cells. Therapy - Booth 720 Yuki Arimoto, New Business Development, Audrey Le, Association Project Manager Medical Devices Huron Technologies - Booth 1002 375 W. 5th Ave. 2-5-8 Higashi-Shinagawa Jeremy Thiel, Sales Manager Vancouver, BC V5Y 1J6 Shinagawa-Ku 550 Parkside Drive, Unit B6 Canada Tokyo, 140-0002 Waterloo, ON N2L 5V4 Phone: +1-604-874-4366 Japan Canada Fax: +1-604-874-4378 Phone: +81-3-5461-3152 Phone: +1-519-886-9013 x39 [email protected] Fax: +81-3-5461-3074 Fax: +1-519-886-5300 www.celltherapysociety.org [email protected] [email protected] ISCT is a global organization that supports pre- www.huron-technologies.com KANEKA’s Bone Marrow MSC Separation Device clinical research, clinical translation, regulatory harvests mesenchymal stem cells (MSC) with a Huron Technologies develops world leading and commercial development of cell-therapy- high recovery of MSCs in a short time (approx. 20 slide scanning solutions for digital pathology. based applications. min.); in a closed system. Also displaying the table The TISSUEscope™ is a high resolution versatile ISCT: top type Automatic Culture System for MSCs. platform capable of imaging specimens up to 8” • Foster networking and educational x 6” in confocal fluorescence and brightfield. We opportunities also offer a scanning service and custom designed • Support the transformation of research into instruments. products and practice

49 ISSCR 9th Annual Meeting www.isscr.org

Exhibitor/Supporter Directory

Kawasaki Heavy Industries Ltd - Leica Microsystems - Booth 630 Mary Ann Liebert, Inc - Booth 433 1700 Leider Lane Booth 713 Yu Iwamoto, Assistant Manager, Buffalo Grove, IL 60089 Lisa Pierce, Exhibits Manager & Society International Business Dept., Corporate USA Marketing Liaison Business Development Div. Phone: +1-800-248-0123 140 Huguenot St. Hamamatsu-cho 2-4-1, Minato-ku Fax: +1-847-405-0164 New Rochelle, NY 10801 Tokyo, 105-6116 [email protected] USA Japan Leica Microsystems will display the Leica DMI Phone: +1-914-740-2100 Phone: +81-3-3435-2175 LED Inverted Digital Microscope, designed to Fax: +1-914-740-2101 Fax: +81-3-3435-2024 provide the best and easiest imaging, every time [email protected] or [email protected] for every user. We will also show the Leica M205 [email protected] www.khi.co.jp/english/ stereomicroscope with Fusion Optics technology, www.liebertpub.com which provides high resolution and high depth The automated cell processing of human induced of field simultaneously throughout the entire Mary Ann Liebert, Inc. publishes authoritative pluripotent stem cells (hiPS cells) in large amounts 20:1 zoom range. Come and see Leica’s imaging peer-reviewed journals in new and promising for industrial use. solutions. areas of science and biomedical research including Stem Cells and Development and Cellular LABS, Inc - Booth 603 Life Technologies - Booth 626 Reprogramming. Stop by Booth #713 - review our collection of cutting edge titles! Chad Ronholdt, Director of Business 5791 Van Allen Way Development or Rachel Garcia, Marketing/ Carlsbad, CA 92008 Massachusetts General Hospital Communications USA 6933 B S. Revere Parkway Phone: +1-760-603-7200 Center for Regenerative Medicine Centennial, CO 80112 Fax: +1-760-602-6500 David Machon, Executive Director USA www.lifetechnologies.com 185 Cambridge St., 4th Floor Phone: +1-800-321-6088 Boston, MA 02114 Life Technologies is a global biotechnology USA Silver Fax: +1-303-343-6666 tools company dedicated to improving the Phone: +1-617-643-5380 [email protected] or rachel_garcia@ human condition. Our customers do their work Supporter labs-inc.org across the biological spectrum, working to Fax: +1-617-724-2662 www.labs-inc.org advance personalized medicine, regenerative [email protected] science, molecular diagnostics, agricultural www.massgeneral.org/regenmed At LABS, Inc. we are committed to helping and environmental research, and 21st century our clients develop innovative cell based The CRM is dedicated to understanding how forensics. products and therapies for use in basic research, tissues are formed and repaired. The goal of our tissue engineering and regenerative medicine diverse, cohesive team of scientists and clinicians applications. We leverage our expertise in Lonza - Booths 701, 703 is to develop novel therapies to regenerate the biological sciences with our dedication to Don Paul Kovarcik, Product Manager, Drug damaged tissues and overcome debilitating delivering accurate, timely and reliable results. Discovery chronic disease. LABS, Inc. is your ideal partner to move your idea 8830 Biggs Ford Road from concept and development through scale-up Walkersville, MD 21793 Silver Massachusetts Human Stem Cell and commercialization. USA Supporter Bank & International Registry - Phone: +1-800-521-0390 Booth 722 Lieber Institute for Brain [email protected] Amy Briggs, Relations Manager/Training Development www.lonza.com Coordinator Ronald McKay, PhD, Director for Basic Lonza supports stem cell and molecular biology 222 Maple Ave Science research with novel tools; Clonetics™ and Shrewsbury, MA 01545 855 N. Wolfe St., Third Floor Poietics™ Primary Cells and Media; Amaxa™ USA Baltimore, MD 21205 Nucleofector™ Transfection; BioWhittaker™ Phone: +1-508-856-7744 USA Silver Media and Sera; SeaKem®, MetaPhor™, Fax: +1-508-856-6566 Phone: +1-301-312-2824 NuSieve™ Agarose; FlashGel™ System, PAGEr™ [email protected] Supporter Precast Protein Gels, and BioAssay Products and [email protected] www.umassmed.edu/iscr www.lieberinstitute.org Services. The MA Stem Cell Bank provides the research The Lieber Institute for Brain Development is community with banking and distribution of a private, non-profit research institute whose expertly maintained and characterized hES mission is to develop new insights into the and iPS cell lines. The International Registry is etiology of behavioral disorders stemming from a searchable and comprehensive database of abnormalities in human brain development information on more than 1200 hES and iPS cell leading to novel therapeutic targets for lines. schizophrenia and related neurodevelopmental disorders.

50 www.isscr.org Final Program

Exhibitor/Supporter Directory

Matrigen - Booth 1018 Molecular Devices, Inc - National Center for Regenerative Justin Mih, Chief Scientific Officer Booth 800 Medicine – Booth 739 60 Prescott St #123 Susan Murphy, Director of Sales Elizabeth Sump Worcester, MA 01605 1311 Orleans Drive 10900 Euclid Ave, LC 7284 USA Sunnyvale, CA 94089 Cleveland, OH 44106 Phone: +1-847-492-3712 USA USA [email protected] Phone: +1-408-747-3633 Phone: +1-216-368-2079 www.matrigen.com Fax: +1-408-747-3601 Fax: +1-216-368-6020 Matrigen is a startup manufacturer of cell culture [email protected] [email protected] platforms based in Worcester, MA. Matrigen’s www.moleculardevices.com www.ncrm.us mission is to innovate and bring to market more At Molecular Devices, understanding your The National Center for Regenerative Medicine physiologically relevant cell culture platforms with laboratory workflow is our top priority, and we has leading research and clinical programs in an emphasis on simplicity, cost effectiveness and direct product development toward solving heart disease, cancer, genetic disorders, wound high-throughput capability. your unique issues. Our instruments offer a full healing and immunological, musculoskeletal and spectrum of detection technologies and meet all neurodegenerative diseases. A 26-year history of McEwen Centre for Regenerative throughput needs to detect biology, decode data research on adult and pluripotent stem cells, and Medicine - Booth 801 and drive discovery. Stop by booth #800, or visit outstanding clinical and research programs makes Heidi Forman, Director of Operations and www.MolecularDevices.com. this Center unique in the United States. Marketing MaRS Centre/TMDT 8-701 MP Biomedicals - Booth 331 National Institute of Biomedical 101 College St. ISSCR John Euston, Strategic Sales Director Imaging and Bioengineering - Toronto, ON M5G 1L7 9th Annual 3 Hutton Centre Drive, Suite 00 Booth 711 Canada Meeting Santa Ana, CA 92707 Shirley Coney-Johnson, Program Analyst, Phone: +1-416-581-7526 Co-Sponsor USA Office of Science Policy and Public Liaison Fax: +1-416-595-5719 Phone: +1-559-259-7798 31 Center Drive [email protected] Fax: +1-800-334-6999 Rm 1C28, MSC 2281 www.mcewencentre.com [email protected] Bethesda, MD 20892 www.mpbio.com The McEwen Centre supports translational USA research that is focused on using stem cells to MP Biomedicals - New Zealand provides bovine- Phone: +1-301-596-9208 accelerate cures and develop more effective derived products with the highest possible Fax: (301) 480-1613 treatments for diseases and conditions including biosafety level for research and industrial [email protected] heart disease, diseases of the blood, diabetes, applications with built-in quality and complete www.nibib.nih.gov respiratory and neurodegenerative diseases. traceability. The portfolio includes bovine albumins, transferrins and fetal bovine serum with The National Institute of Biomedical Imaging and Bioengineering leads development Miltenyi Biotec GmbH - demonstrated success in culturing stem cells and improving protein production from primary cell and application of breakthrough biomedical Booth 418 lines. technologies in order to improve human Dr. Dirk Winnemöller, Global Marketing health. NIBIB is committed to integrating the Manager Stem Cells MVE Chart - Booth 805 engineering, physical and life sciences to advance basic research and medical care. Friedrich-Ebert-Str 68 Bruce Edel, Sales Manager Bergisch Gladbach, 51429 2200 Airport Industrial Drive, Suite 500 Nature Publishing Group – Germany Ball Ground, GA 30107 Booth 631 Phone: +49-2204-83060 USA 75 Varick St, 9th floor Fax: +49-2204-85197 Phone: +1-770-721-7700 New York, NY 10013 [email protected] Fax: +1-770-721-7701 USA www.miltenybiotec.com [email protected] Phone: +1-212-726-9200 Miltenyi Biotec has provided cutting-edge www.chartbiomed.com Fax: +1-212-696-9006 scientific tools to support stem cell research MVE Chart features a complete line of aluminum www.nature.com for more than 15 years. The portfolio includes vapor shippers and nitrogen handling equipment. reagents for fundamental research through to We introduce the energy-efficient Vario Freezer The NPG portfolio combines the excellence of clinical applications and covering adult stem cells into our line of liquid and vapor stainless steel Nature, its associated research, review journals, as well as ES and iPS cells. freezers with 3,200-94,000 vial capacities. leading academic and society journals in the life, physical and clinical sciences. Open access options are offered through the academic and society journals, Nature Communications.

51 ISSCR 9th Annual Meeting www.isscr.org

Exhibitor/Supporter Directory

The New York Stem Cell Norgen Biotek Corp. - Booth 928 Ontario Stem Cell Initiative (OSCI) Foundation (NYSCF) - Booth 719 Tom Hunter, Marketing Manager - Booth 710 Kacey Koeppel, Senior Director of Events 3430 Schmon Parkway Sandra Donaldson, Program Manager 1995 Broadway, Suite 600 Thorold, ON L2V 4Y6 1110 - 160 College St New York, NY 10023 Canada Toronto, ON M5S 3E1 Bronze USA Silver Phone: +1-905-227-8848 Canada Phone: +1-212-365-7452 Supporter Fax: +1-905-227-1061 Phone: +1-416-978-0553 Supporter Fax: +1-212-787-5844 [email protected] Fax: +1-416-978-2666 [email protected] www.norgenbiotek.com [email protected] www.nyscf.org Norgen Biotek is an innovative biotechnology www.ontariostemcell.ca NYSCF is a non-profit organization dedicated company specializing in sample preparation The Ontario Stem Cell Initiative is a virtual to accelerating stem cell research through its technologies. We produce novel RNA, DNA, network of more than 60 stem cell scientists in independant research laboratory, collaborations, protein and multiple-analyte purification, isolation Ontario. The Centre for the Commercialization grants, scientific conferences and public and clean-up kits based on our proprietary of Regenerative Medicine is a not-for-profit education. technology. Norgen’s unique RNA Purification organization dedicated to the development Kits isolate all sizes of RNA, including microRNA, of technologies that accelerate the without the use of phenol and with excellent Nexcelom Bioscience - Booth 925 commercialization of stem cell- and biomaterials- sensitivity. based technologies and therapies. Lyndsey Vavra, Sales Operations Manager 360 Merrimack St Novus Biologicals - Booth 422 ORIGIO, Inc - Booth 820 Lawrence, MA 01843 Kim Mesman, Marketing Manager Mary Molinari, Sales Representative USA 8400 Southpark Way, A-8 77 Elbo Lane Phone: +1-978-327-5340 Littleton, CO 80120 Mount Laurel, NJ 08054 Fax: +1-978-327-5341 USA USA [email protected] Phone: +1-303-730-1950 Phone: +1-800-648-1151 www.nexcelom.com Fax: +1-303-730-1966 Fax: +1-856-762-2009 Nexcelom’s Cellometer line of simple-to-use [email protected] [email protected] cell counters automate manual cell counting www.novusbio.com www.origio.com procedures by obtaining accurate counts, viability and cell sizes in less than 30 seconds & only 20uL Novus Biologicals provides stem cell research tools Conceived from the specialized building blocks of sample. Fluorescence detection capabilities including antibodies, stem cell lines, proteins, of MediCult, MidAtlantic Diagnostics and enable fast & simple determination of GFP peptides, isotype controls, loading controls and Humagen. ORIGIO offers a one-stop solution for transfection rates, PI-viability, & direct counting more. All products are 100% guaranteed to any research lab. Products range from disposable of WBCs without lysing. work in the species and applications listed on the microtools to media to specialized equipment to a datasheet. Visit www.novusbio.com to browse revolutionary air purification system. Novus’ stem cell catalog. NOF America Corporation - Booth 924 Paragon Bioservices, Inc - Olympus Canada, Inc - Booth 431 Booth 1012 Hirofumi Irie, Manager, Life Science Roxanne DeAbreu-Breen, Marketing Marco A Chacon, PhD, President & CEO Products Coordinator 801 W Baltimore St, Suite 401 One North Broadway, Suite 1012 151 Telson Road Baltimore, MD 21201 White Plains, NY 10601 Markham, ON L3R 1E7 USA USA Canada Phone: +1-410-975-4050 Phone: +1-914-681-9790 Phone: +1-905-969-3201 Fax: +1-410-605-2028 Fax: +1-914-681-9791 Fax: +1-905-479-2595 [email protected] [email protected] [email protected] www.paragonbioservices.com www.nofamerica.com www.olympuscanada.com A contract research and manufacturing We, NOF AMERICA CORPORATION, are Olympus Canada Scientific Equipment Group a US subsidiary of NOF CORPORATION, a organization (CMO) that serves as a translational provides innovative microscope and image bridge, enabling technologies to go from the chemical company based in Japan. We are analysis solutions to researchers, doctors, providing Lipidure-Coat, low cell binding vessels research lab to Phase I/II clinical trials. Our clinicians, educators as well as to most industrial areas of expertise include GMP manufacturing coated with biocompatible polymer containing market sectors. Olympus microscope systems phosphorylcholine moiety in its structure. of monoclonal antibodies, therapeutic proteins, offer unsurpassed optics, superior construction vaccines (VLPs) and stem cells. Therefore, spherical embryoid bodies can be and system versatility to meet the ever-changing obtained thanks to high biocompatibility. needs of microscopists, paving the way for future advances in life science.

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Exhibitor/Supporter Directory

PeproTech, Inc - Booth 526 profiling of whole genomes and miRNA. Our QIAGEN, Inc - Booth 709 Heather Giordano, Marketing Coordinator highly-trained specialists provide comprehensive Pam Daniels, Events Manager 5 Crescent Ave offerings for gene expression and miRNA profiling 19300 Germantown Road services including RNA extraction, microarray Rocky Hill, NJ 08553-0275 Germantown, MD 20874 hybridization, qPCR and analysis consultation. USA USA Phone: +1-800-436-9910 Phone: +1-240-686-7688 Primorigen Biosciences, Inc - Fax: +1-609-497-0321 Fax: +1-240-686-7689 [email protected] Booth 705 [email protected] www.peprotech.com Bradley H. Garcia II, PhD, Director of www.qiagen.com Technology and Business Development PeproTech manufactures an extensive line of 510 Charmany Drive QIAGEN Inc., the worldwide leading provider recombinant human, murine and rat proteins, of sample and assay technologies for research Madison, WI 53719 animal-free recombinant proteins, monoclonal in life sciences, applied testing and molecular antibodies, affinity purified polyclonal antibodies, USA diagnostics, offers more than 500 proprietary affinity purified biotinylated polyclonal antibodies, Phone: +1-608-628-5189 products sold through a dedicated sales force and ELISA development kits and serum-free media Fax: +1-608-441-8333 a global network of distributors. supplements. [email protected] www.primorigen.com R&D Systems, Inc - Booth 914 Pfizer Canada Inc Primorigen Biosciences develops innovative Tracey Husted, Tradeshow Coordinator Dr. Mark Lundie solutions for stem cell proliferation, 614 McKinley Place NE 17300 Trans Canada Highway characterization and assay development, Minneapolis, MN 55413 Kirkland, QC H9J 2M5 including defined substrates for culturing, large USA Canada Silver scale cell selection and production systems, Phone: +1-612-379-2956 multiplexed immunoassays for stem cell protein Phone: +1-514-695-0500 Fax: +1-612-379-6580 Supporter profiling, and stem-cell derived assays for [email protected] compound screening. [email protected] www.pfizer.ca www.rndsystems.com Pfizer Canada Inc. is the Canadian operation of PromoCell GmbH - Booth 1004 R&D Systems serves basic and clinical researchers Pfizer Inc, the world’s leading biopharmaceutical Dr. Lorna Whyte, Market Development with products for cell biology research. We company. Pfizer discovers, develops, Mananager provide purified proteins; immunohistochemistry manufactures and markets prescription medicines Sickingenstrasse 63/65 and flow cytometry-validated antibodies; for humans and animals. At Pfizer, we’re working ELISA kits, including multiplex formats, to a Heidelberg, D-69126 together for a healthier world. variety of targets for studying cellular growth, Phone: +1-866-251-2860 differentiation and function. Pfizer Neusentis Fax: +1-866-827-9219 Dr. Timothy Allsopp, Head of External [email protected] ReachBio - Booth 601 www.promocell.com Research Rob Chaney, Operations Director PromoCell - The Cell Culture Experts The Portway Building, Granta Park 454 N 34th St Cambridge CB21 6GS At PromoCell, we are committed to providing Seattle, WA 98103 United Kingdom Silver researchers worldwide with a broad range of USA Phone: +44-1304-643483 human primary cells, stem and blood cells, Phone: +1-206-420-0300 optimized cell culture media as well as growth [email protected] Supporter Fax: +1-206-420-0301 factors and cytokines for stem cell research. Neusentis is a novel business unit [email protected] incorporating regenerative medicine, pain Proteintech Group Inc - www.reachbio.com & sensory disorders & epigenetics based in ReachBio provides primary cell-based products Cambridge, U.K. Booth 319 Katie Strong, Customer Service Director and contract assay services. We focus in blood and bone marrow systems offering specialized 2201 W Campbell Park Drive Phalanx Biotech Group - media for hematopoietic (ColonyGEL™) and Chicago, IL 60612 mesenchymal progenitor stem cell culture and Booth 815 USA various primary cell (REACHBIO QUALIFIED™) Kevin Poon, Technical Marketing and Phone: +1-312-455-8498 products. Support Manager Fax: +1-312-455-8408 1301 Shoreway Road, Suite 160 [email protected] Belmont, CA 94002 www.ptglab.com USA Proteintech’s mission is to fulfill Human EST fusion Phone: +1-650-320-8669 protein Project and Proteomic Antibody Project. Fax: +1-650-508-9889 We’ve produced more than 10,000 high quality [email protected] or antibodies validated by WB and IHC on primary [email protected] tissues and cell lysates. Worldwide locations, www.onearray.com antibodies in stock, next day delivery. Phalanx Biotech offers the OneArray® line of high-quality, affordable microarrays for expression 53 ISSCR 9th Annual Meeting www.isscr.org

Exhibitor/Supporter Directory

Regenerative Medicine, Future Roche Diagnostics Corporation - Scottish Development Medicine - Booth 804 Booth 932 International - Booth 823 Craig Canham, Marketing Manager Alexander R. Domingo, Account Manager Joanna Gourlay or Larry Reynolds Unitec House, 2 Albert Place 9115 Hague Road 5 Atlantic Way, 150 Broomielaw London, N3 1QB Indianapolis, IN 46250 Glasgow, G2 8LU United Kingdom USA United Kingdom Phone: +44-20-8371-6088 Phone: +1-317-521-1922 Phone: +44-141-8423524 Fax: +44-20-8371-6089 Fax: +1-317-521-4065 [email protected] or larry. [email protected] [email protected] [email protected] www.futuremedicine.com www.collagenase.com www.sdi.co.uk Regenerative Medicine is the award winning Roche is setting a new standard for adipose tissue Scotland is a world leader in stem cell and forum for a fast-growing community of specialists dissociation with the introduction of a Sterile-A regenerative medicine R&D, both academic and to address the important challenges and advances “mammalian tissue-free” Liberase Enzyme. This commercial. Scottish Development International’s that are now occurring in stem cell research and highly purified Collagenase blend offers a tool for aim is to assist in the growth of the Scottish regenerative medicine. optimizing stem-cell isolations to maximize cell economy, by encouraging inward investment yield, viability, functionality and safety. and helping Scottish-based companies develop ReproCELL - Booth 829 international trade. Hiroko Nakamura, Marketing Manager SANYO - Booth 910 560 S Winchester Blvd, Suite 500 Deepak M. Mistry, Manager, Strategic Sigma Life Science - Booth 903 San Jose, CA 95128 Development & Marketing Jennifer Williams, Product Manager USA 1300 Michael Drive, Suite A 3050 Spruce St Phone: +1-408-876-8544 Wood Dale, IL 60191 Saint Louis, MO 63103 Fax: +1-408-572-5679 USA USA [email protected] Phone: +1-800-858-8442 Phone: +1-314-771-5765 www.reprocell.net Fax: +1-630-238-0074 [email protected] [email protected] www.wherebiobegins.com/ ReproCELL provides products and services for bioreprogramming basic research and drug discovery, based on http://us.sanyo.com/biomedical leading stem cell technologies developed in Biomedical Solutions, a Division of SANYO North SIGMA® Life Science offers a wide portfolio of Japanese institutes. Products include cell culture America, is based in Wood Dale, IL, and provides innovative technologies, products and services reagents and antibodies for human ES/iPS-cell laboratory equipment for pharmaceutical, life spanning cell biology, protein assays, genomics, research, hiPS cell-derived cardiomyocytes and science and biotechnology applications. Product functional genomics, biomolecules, epigenetics, neurons, with hepatocytes forthcoming. lines include space-efficient VIP® ultra-low stem cell research and transgenic animal models. temperature freezers, cryogenic freezers, Through continued investment in innovation RESBIO-NIH Funded Biomedical pharmaceutical and medical refrigerators, cell and quality we are the leading destination for life science researchers to access deep biological Technology Resource - Booth 629 culture incubators, plant growth chambers and portable autoclaves. information and market leading products that Pallassana Narayanan, PhD, Chief improve the quality of life. Operating Officer Sarstedt Inc - Booth 605 145 Bevier Road Alain Cote, Manager Sales or Eric Simoneau, Society for Neuroscience - Piscataway, NJ 08854 Assisant Sales Manager Booth 821 USA 6373 Des Grandes Prairies Wendy Sturley, Director, Membership and Phone: +1-732-445-0488 Montreal, QC H1P 1A5 Marketing Fax: +1-732-445-5006 Canada 1121 14th St, NW, Suite 1010 [email protected] Phone: +1-888-727-7833 Washington, DC 20005 www.njbiomaterials.org Fax: +1-514-328-9391 USA RESBIO is an NIH-funded national biomedical [email protected] Phone: +1-202-962-4000 technology resource that develops integrated www.sarstedt.com Fax: +1-202-962-4946 multidisciplinary state-of-the-art technologies [email protected] tailored to accelerate biomaterials research. Sarstedt has a complete line of cell culture www.sfn.org The booth will showcase high-throughput products with plates, dishes, flasks, also reusable polymeric biomaterial synthesis, high-throughput silicone inserts and lumox dishes, plus a wide Founded in 1969 with 500 members, the Society cell material interaction characterization, range of cryovials. for Neuroscience has grown to more than 41,000 mechanotransduction research, confocal in 87 countries. Stop by our booth to learn more microscopy for early determination of stem about our annual meeting, Neuroscience 2011, cell lineage commitment, and computational taking place November 12-16 in Washington modeling for biomaterial optimization. D.C. or pick up a copy of The Journal of Neuroscience.

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Exhibitor/Supporter Directory

Springer - Booth 728 STEMCELL Technologies Inc - Sutter Instrument - Booth 408 Acasia Dalmau, Exhibits Manager - USA Booth 506 Alexandra Cooper, Sales And Marketing 233 Spring St Clive Glover, Manager, Product One Digital Drive New York, NY 10013 Management Novato, CA 94949 USA 570 West 7th Ave., Suite 400 USA Phone: +1-212-460-1500 ext. 1600 Vancouver, BC V5Z 1B3 Phone: +1-415-883-0128 Fax: +1-201-460-1575 Canada Fax: +1-415-883-0572 [email protected] Phone: +1-604-877-0713 Silver [email protected] www.springer.com Fax: +1-604-877-0704 Supporter www.sutter.com Get hands-on experience with Springer’s [email protected] Sutter Instrument is a leading manufacturer of multi-format publishing model: print – eBook – www.stemcell.com scientific instrumentation. The XenoWorks® MyCopy (printed eBooks for 24.95 EUR/USD). STEMCELL Technologies is a leading provider microinjection system is designed for a wide With more than 3,400 books currently available in of specialized cell culture media for pluripotent, variety of applications (ICSI, ES cell, pronuclear the Biomedical & Life Sciences eBook Collection, mesenchymal, neural, mammary epithelial and injection, etc.). Our bevelers and programmable our mission is to support your research. Come hematopoietic stem cell research. A full line of micropipette pullers are ideal the fabrication of browse our books in your preferred format: cell separation products, including RoboSep™, precision microtools. print, online, or on an eReader or iPad. Ensure the fully automated cell separator, are available optimized print and electronic dissemination of for the isolation of any cell type from virtually System Biosciences (SBI) - your work, too! Get Read. Publish With Springer. any species. Other products and services include Booth 831 primary human cells, sera, cytokines, antibodies, Enal Razvi, PhD, Vice President Of Sales Stem Cell Network - training courses, proficiency testing, and contract 265 N Whisman Road services. Booths 623, 625 Mountain View, CA 94043 Lisa Willemse, Director of Communications Stemgent - Booth 611 USA 501 Smyth Road, Suite CCW-6189 Phone: +1-650-968-2200 Rebecca Rossi, Marketing Communications Ottawa, ON K1H 8L6 Fax: +1-650-968-2277 Specialist Canada [email protected] Diamond One Kendall Square, Suite B6201 Phone: +1-613-739-6675 www.systembio.com Fax: +1-613-739-6680 Supporter Cambridge, MA 02139 [email protected] USA System Biosciences (SBI) develops and Platinum commercializes tools for stem cell research and www.stemcellnetwork.ca Phone: +1-877-228-9783 Fax: +1-617-494-0398 Supporter microRNA research. Our broad portfolio of The Stem Cell Network is a not-for-profit products for stem cell research includes viral [email protected] organization, supporting leading-edge research and minicircle-based reprogramming factors for and training at more than 30 institutions across www.stemgent.com generating iPSCs, protein-derived iPSC lines, stem Canada, with a goal to catalyze the translation Stemgent has been working alongside some of cell differentiation reporters, as well as premade of stem cell research into clinical applications, the world’s leading stem cell scientists to develop iPSC lines-both normal and disease-specific. We commercial products and public policy. stem cell-qualified reagents including small have a growing portfolio of disease-specific iPSCs molecules, media, tools for the generation of iPS including diabetes, ALS, SMA, MLD and others. StemCells Inc cells and more for the advancement of stem cell Additionally, SBI offers custom reprogramming services-either using viral-based reprogramming Stewart Craig, PhD, Sr. VP Development & research. methods or minicircle-based reprogramming. Operations 3155 Porter Drive StemRD Inc - Booth 708 Silver TAP Biosystems - Booth 427 Palo Alto, CA 94304 Songzu An, President & CSO Matthew Walker, Head of Communications USA Supporter 332 Beach Road 20 Monchanin Road Phone: +1-650-475-3100 Burlingame, CA 94010 Greenville, DE 19807 Fax: +1-650-475-3101 USA USA [email protected] Phone: +1-650-343-1888 Phone: +1-302-478-9060 www.stemcellsinc.com Fax: +1-650-343-1889 [email protected] Fax: +1-302-475-9575 Research, development and commercialization www.stemrd.com [email protected] of cell-based therapeutics and tools for use in www.tapbiosystems.com stem cell-based research and drug discovery. StemRD, Inc. manufactures premier stem cell Our HuCNS-SC® human neural stem cells are in research reagents using proprietary technology TAP Biosystems (formerly The Automation clinical development for CNS disorders. We also to efficiently produce highly pure, highly active Partnership) provides advanced automation market SC Proven® media and reagents and stem WNT, Hedgehog, TGF-beta and other factors, systems and services to improve productivity in cell-based assay platforms. often in human cells. Exclusive products include stem cell research applications. TAP will showcase human WNT-5a and Mesengro® chemically- the CellBase CT for automated ex vivo expansion defined mesenchymal stem cell medium. of autologous cell therapy products, plus the Cell- IQ system for studying live cell behavior.

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Exhibitor/Supporter Directory

Taylor Wharton - Booth 309 TOKAI HIT Co., Ltd - Booth 809 University of Pittsburgh, Jim Gordon, CryoScience Technical Director Aki Doi, Global Sales Manager McGowan Institute for 4075 Hamilton Blvd 306-1 Gendoji-cho Regenerative Medicine Theodore, AL 36582 Fujinomiya-City, 418-0074 Alan Russell, PhD, Director USA Japan 450 Technology Drive, Suite 300 Phone: +1-717-443-0750 Phone: +81-544-24-6699 Pittsburgh, PA 15291 Fax: +1-717-620-3311 Fax: +81-544-24-6641 USA Bronze [email protected] [email protected] Phone: +1-412-624-5500 www.taylorwharton.com www.tokaihit.com/english/top/index.html Fax: +1-412-624-5363 Supporter Taylor-Wharton’s is an industry leader in the For live-cell-imaging. Tokai Hit presents Stage [email protected] design of cryogenic products and inventory Top Incubator INU-series and Gas-Mixer for [email protected] systems. Taylor-Wharton’s CryoScience products Hypoxia. INU provides optimal tissue/cell culture www.mcgowan.pitt.edu include; aluminum refrigerators, dry shippers, environment on microscope-stage that enables dewars and -190C high efficiency vapor freezers long-term-timelapse to obtain continuous images The McGowan Institute serves as a single base storing in excess of 94,000 vials. of migration, differentiation, propagation etc. of operations for the University of Pittsburgh of stem/ES/iPS cells. Clear-Glass-Heater is also scientists and clinical faculty working to develop available. tissue engineering, cellular therapies and artificial Thermo Scientific - and biohybrid organ devices. The Institute’s Booths 830, 832 mission includes the development of innovative Jessica Weiss, Trade Show Manager Tree Star, Inc - Booth 812 clinical protocols as well as the pursuit of rapid 2650 Crescent Drive Suzanne Mertens, Application Scientist commercial transfer of its technologies. The focus Lafayette, CO 80026 340 A Street is on the repair or replacement of tissues and organs by the use of bioengineered materials, USA Silver Ashland, OR 97520 cells, genes or other biological building blocks. Phone: +1-303-604-3224 Supporter USA Fax: +1-303-604-3289 Phone: +1-800-366-6045 VisualSonics Inc - Booth 723 [email protected] Fax: +1-541-482-3153 Mandi Waite, Marketing Coordinator www.thermoscientific.com/stemcell [email protected] www.flowjo.com 3080 Yonge St, Suite 6100 Come share your most demanding stem cell Toronto, ON M4N 3N1 FlowJo is the next generation of flow cytometry challenges. Talk one-on-one with technical Canada experts. Visit our booth to learn about the latest analysis software. It handles your most ambitious Phone: +1-416-484-5000 advances in personal imaging cytometry, specialty projects with a high-level drag-and-drop user surfaces for cell culture, molecular biology tools interface. Based on a patented experiment-based Fax: +1-416-484-5001 and more. analysis paradigm, FlowJo intelligently handles [email protected] protocols containing multiple tubes (any FCS www.visualsonics.com files). Tissue Regeneration Therapeutics VisualSonics is the undisputed world leader in real-time, in vivo, high-resolution, micro- Inc - Booth 730 Union Biometrica, Inc - Booth 501 Zeesy Powers, Office Manager and imaging systems, providing modalities specifically Michael Fazzio, Director of Sales, Northeast designed for preclinical research. These Research Assistant 84 October Hill Road cutting edge technologies allow researchers 790 Bay St., Suite 512 Holliston, MA 01746 to conduct research in cardiovascular, cancer Toronto, ON M5G 1N8 USA and neurobiology areas. VisualSonics platforms Canada Phone: +1-973-610-8190 combine high-resolution, real-time in vivo Phone: +1-416-345-0039 imaging at a reasonable cost with ease-of-use Fax: +1-508-893-8044 Fax: +1-416-345-9277 and quantifiable results. [email protected] [email protected] www.unionbio.com www.verypowerfulbiology.com Vitrolife - Booth 702 Union Biometrica Large Particle Flow Cytometers Anna Mårtensson, Business Area Manager Tissue Regeneration Therapeutics extracts automate the analysis, sorting and dispensing of perivascular cells from umbilical cord, Human Gustaf Werners Gata 2 objects too big/fragile for traditional cytometers, Goteborg, 40092 Umbilical Cord PeriVascular Cells (HUCPVCs). e.g., large cells / cell clusters, cells in/on beads Sweden The high CFU-F frequency within HUCPVCs has and small . Choose between provided the first robust experimental proof of 4 COPAS models and the new BioSorter with Phone: +46-708-22-8029 the existence of MSC, and they have increased interchangeable modules to cover the full 10- Fax: +46-31-721-8099 biological potential compared cells derived from 1500um range. [email protected] adult tissues. www.vitrolife.com Vitrolife is an international biotechnology/ medical device group. We develop, manufacture and sell advanced products and systems for the preparation, cultivation and storage of human cells, tissue and organs.

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Exhibitor/Supporter Directory

WaferGen Biosystems, Inc - Wiley Blackwell - Booth 818 Booth 329 Virginia Chanda Paul Kotturi, Sr. Product Manager 111 River St. 7400 Paseo Padre Parkway Hoboken, NJ 07030 Silver Fremont, CA 94555 USA Supporter USA Phone: +1-203-748-6000 Phone: +1-510-651-4450 [email protected] Fax: +1-510-277-3420 www.wiley.com [email protected] Wiley-Blackwell is the international scientific, www.wafergen.com technical, medical and scholarly publishing WaferGen Biosystems offers the SmartChip business of John Wiley & Sons, with strengths in System, which combines the high-throughput every major academic and professional field and of microarrays with the power of real-time PCR partnerships with many of the world’s leading to provide the unique flexibility to perform gene societies. For more information, please visit expression discovery, validation and screening on www.wileyblackwell.com or a single platform. http://onlinelibrary.wiley.com/

WiCell Research Institute - Worthington Biochemical Booth 715 Corporation - Booth 900 Robert J. Drape, Director of Operations Jim Zacka, VP 505 Rosa Road, Suite 120 730 Vassar Ave Madison, WI 53719 Lakewood, NJ 08701 USA USA Phone: +1-608-441-8025 Phone: +1-800-445-9603 or Fax: +1-608-441-8011 +1-732-942-1660 [email protected] Fax: +1-800-368-3108 or www.wicell.org +1-732-942-9270 [email protected] WiCell Research Institute is a nonprofit www.worthington-biochem.com organization established in 1999 to advance stem cell technology. WiCell distributes pluripotent Worthington produces Animal Origin Free stem cells to researchers worldwide and offers Collagenase and enzymes for regenerative cytogenetic services, training courses, testing medicine applications. Proteases, nucleases and services and research & clinical grade cell lines. kits for stem cell and primary cell isolation & optimized cell isolation kits. Request our NEW Tissue Dissociation Guide or visit our Website at www.worthington-biochem.com

57 58 www.isscr.org Final Program

Detailed Program and Abstracts – Wednesday, June 15

Plenary I – Presidential Symposium: REGENERATIVE MEDICINE: TISSUE ENGINEERING, BIOMATERIALS AND Stem Cell Biology and the Prospects for Regenerative CONTROLLED DRUG DELIVERY Medicine: Past, Present and Future Langer, Robert Supported by COSAT – Johnson & Johnson Dept of Chemical Engineering, Massachusetts Institute of Technology, Cambridge, MA, USA Wednesday, June 15, 1:00 – 3:30 p.m. By combining mammalian cells, including stem cells, with synthetic poly- FROM TILL AND MCCULLOCH TO IPS CELLS — mers, new approaches for engineering tissues are being developed that may someday help repair tissues for patients with burns, damaged cartilage, THE IMPORTANCE OF FUNCTIONAL ASSAYS paralysis and vascular disease. Various technological advances in controlled Rossant, Janet drug delivery, nanotechnology and other areas may aid in the development of these approaches. These advances include controlled delivery of bioac- Developmental and Stem Cell Biology, Hospital for Sick Children, Toronto, tive agents including DNA and siRNAs, the development of new biomateri- ON, Canada als and the creation of new materials-based tools for understanding stem 2011 marks the 50th anniversary of the publication by Jim Till and the late cell growth and differentiation. Ernest McCulloch of a functional clonal assay, the ‘spleen colony forming assay’, demonstrating that rare self-renewing cells in the hematopoietic system retained the capacity to differentiate into multiple blood cell types. TITLE TBD Thus they established the stem cell paradigm that underlies all stem cell research today. Stem cells, both multipotent and more restricted, have been Weissman, Irving L. identified in many different tissues and stages of development but not all Stanford University School of Medicine, Palo Alto, CA, USA have been validated by the kind of rigorous clonal assay pioneered by Till and McCulloch. Pluripotent stem cell research traces its origins to the work ABSTRACT NOT AVAILABLE AT TIME OF PRINTING on spontaneous and induced testicular teratocarcinomas in the 1950s and 60s, pioneered by Leroy Stevens and Barry Pierce. Pierce used a clonal transplantation assay to show that a single cell could recapitulate the entire diversity of cell types in the tumor. Not only did this prove that a single Plenary II – Totipotency and Germline Development stem cell could be pluripotent but it was also the first demonstration of a Wednesday, June 15, 4:15 – 6:30 p.m. cancer stem cell. Again a functional clonal assay was key. An even more stringent assay for pluripotency was then proposed— namely the ability PROTECTING THE GERMLINE THROUGH to recapitulate normal development when introduced into the early mouse SILENCING embryo — the chimera assay. Teratocarcinoma stem cells were genetically unstable and not able to recapitulate entirely normal embryonic develop- Lehmann, Ruth, Rangan, Prashanth, Zamparini, Andrea, ment, but when mouse embryonic stem (ES) cells were isolated from the Malone, Colin blastocyst it was soon shown that single ES cells could contribute to all cell Skirball Institute, Kimmel Center for Stem Cell Biology, NYU School of lineages in a chimera. As we move towards human ES and iPS cells, the Medicine, New York, NY, USA development of a ‘gold standard’ functional assay to establish pluripotency has been a major challenge. We need to continue to refine our functional Protecting the genome from transposable element (TE) mobilization is criti- assays with the experimental rigor invoked by the early stem cell pioneers. cal for germline development. In Drosophila, Piwi proteins associate with Piwi-interacting RNAs (piRNAs) and protect the germline genome by silenc- ing TEs. TE targeting piRNAs are processed from TE-dense heterochromatic DIRECTING AND REDIRECTING CELL FATES loci termed ‘piRNA clusters’. Here we discuss how piRNA cluster tran- scription and piRNA biogenesis are regulated in the germline and somatic Daley, George Q. components of the gonad. In the absence of cluster precursor transcription, Children’s Hospital, Boston/HHMI, Harvard Stem Cell Institute, Boston, their processing or loading onto Piwi proteins, TEs are activated in germline MA, USA and somatic gonadal cells resulting in germline loss and a block in germline A major goal of stem cell research is the creation of customized stem cells stem cell differentiation. for use in disease modeling and as a foundation for autologous cell therapy. Understanding the principles of stem cell derivation, and defining whether embryonic stem (ES) cells and induced pluripotent stem (iPS) are equiva- THE RELATIONSHIP BETWEEN SPECIFICATION lent remains a pressing issue. We have derived human and mouse ES cells OF GERM CELLS AND REGULATION OF and iPS cells from various human and mouse tissues, and compared their differentiation capacity and epigenome to determine whether iPS cells PLURIPOTENCY are functional and molecular equivalents of ES cells, and whether iPS cells Surani, Azim retain an “epigenetic memory” of their tissue of origin. Aspects of iPS cell derivation from normal and diseased tissues will be discussed, as will data Wellcome Trust Cancer Research UK Gurdon Institute, University of comparing ES and iPS cells. Cambridge, Cambridge, United Kingdom Primordial germ cells originate from postimplantation epiblast cells in response to BMP4, from cells that have already initiated the process of dif- ferentiation towards somatic cell lineages. Evidence shows that this trend is arrested in epiblast cells that are recruited to form the founder population of germ cells through expression of key germ cell determinants: Blimp1/ Prdm1, Prdm14, Lin28 and Prmt5. The accompanying epigenetic modifica-

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Detailed Program and Abstracts – Wednesday, June 15

tions erase the epigenetic memory of their trajectory towards somatic fate to an immediate alteration in Oct4, Sox2, and Nanog expression. More as early germ cells become epigenetically similar to ICM and ES cells. Addi- specifically, Sox2 expression is downregulated, while Nanog expression is tional epigenetic modifications, including genome-wide DNA demethylation upregulated. Oct4 expression is immediately upregulated, perhaps trigger- and histone modifications in early germ cells are important as they progress ing ES cell differentiation. Our data suggest that Brg1 plays an important towards an epigenetic ground state, and for resetting the epigenetic state of role in regulating Oct4 expression, which is instrumental in maintaining cells germ cells towards the eventual totipotent state. Postimplantation epiblast in a pluripotent state. can also be induced to undergo reversion to ESC-like cells in response to LIF-STAT3 signaling, which results in some similar epigenetic changes as seen during PGC specification, including X reactivation. It is of interest to SPERM STEM CELLS IN THE MOUSE TESTIS understand how mechanistically, epiblast cells acquire two distinct pheno- Yoshida, Shosei typic states. Increasing knowledge of the mammalian germ cell lineage and in vitro epigenetic reprogramming of postimplantation epiblast may provide Division of Germ Cell Biology, National Institute for Basic Biology, Aichi, insights into the mechanisms that regulate their responses to diverse signals, Japan which may induce different epigenetic responses regulating cell fates. In mammalian testis, numerous sperms are produced persistently for a prolonged period, which plays an essential role for the continuity of life. It is generally considered that ‘stem cells’ that are both capable of self-renewal INDUCTION OF PLURIPOTENCY IN GERMLINE and differentiation supports the continuity of spermatogenesis. ‘Stem cells’ are also considered to be crucial not only for steady state spermatogenesis AND SOMATIC CELLS but also for regeneration after damage or transplantation. However, it is still Schöler, Hans1, Arauzo-Bravo, Marcos1, Han, Dong-Wook1, Kim, largely to be elucidated what is the cellular identity and characters of the Jeong-Beom2, Singhal, Nishant1, Tapia, Natalia1, Wu, Guangming1 ‘stem cells’ and how they behave in the testis so that they can give rise to the different aspects of stem cell functions and maintain the spermatogen- 1 Max Planck Institute for Molecular Biomedicine, Muenster, Germany, esis homeostasis. We have been investigating this issue by means of live im- 2 Ulsan National Institute of Science and Technology, UNIST, Ulsan, Korea, aging and pulse labeling, which allow investigating the cellular behavior in Republic of undisturbed testis that undergoes steady-state spermatogenesis. These have The mammalian germline comprises two principal parts: the inner cell mass shown us, slowly but steadily, how the stem cell functions are achieved. and epiblast, containing pluripotent cells, and the germ cell lineage, host- It has been suggested that, rather than a very limited number of defined ing unipotent cells. Several pluripotent stem cell types have been derived ‘stem cells’, extended population of undifferentiated cells with different from the first part. Embryonic stem (ES) cells, derived from preimplantation degree of self-renewing and differentiating abilities compose the functional embryos, comprise at least two populations of cells with divergent states of stem cell compartment. Interestingly, it is largely influenced by the tissue pluripotency. In addition, epiblast stem cells (EpiSCs), derived from postim- situation (steady-state, regenerating after damage, or post-transplantation) plantantion embryos, also do not comprise a uniform cellular population, as how much the cell population is recruited to the active population with the we have shown the presence of at least two EpiSC types: one resembling stem cell functionality. These findings may expand our view regarding the epiblast tissue of early and the other of late pregastrulation embryos. definition of ‘stem cells’ in the mouse spermatogenesis. A current aim in cell and developmental biology is to program cells at will. The first step in converting a given cell into another cell is through achieve- MICRORNAS TO PATHWAYS IN STEM CELL ment of a pluripotent stem cell state that resembles that of ES cells. To date, somatic cells need to be pushed to a pluripotent state by the introduction of FATE DECISIONS exogenous factors, mostly transcription factors. Reprogramming of mouse Blelloch, Robert and human somatic cells into pluripotent stem cells, designated as induced University of California, San Francisco, San Francisco, CA, USA pluripotent stem (iPS) cells, was first described in 2006 using fibroblasts as the somatic cell source and initially requiring introduction of the virally- MicroRNAs are small non-coding RNAs, which individually target and expressed transcription factor quartet of Oct4, Sox2, c-Myc, and Klf4. suppress hundreds of transcripts. Therefore, they are well positioned to As we have recently shown, induced EpiSCs (iEpiSCs) can be obtained by globally regulate cell fate decisions. Indeed, microRNAs alone can promote directly reprogramming somatic cells with the quartet under EpiSC culture the de-differentiation of somatic cells to induced pluripotent stem cells. As conditions. We previously reported that Oct4 alone is sufficient to directly microRNA targets are evolutionarily selected to have specific cellular out- reprogram adult mouse and human fetal neural stem cells (NSCs) into iPS comes, we hypothesized we could use miRNAs to uncover pathways impor- cells, thus highlighting the crucial role played by Oct4 in the process of tant in promoting cell fate decisions. To test this hypothesis, we have begun reprogramming. In contrast to the recent reprogramming of somatic cells, systematically dissecting all the targets of the ESCC miRNAs, which promote induction of pluripotency in primordial germ cells (PGCs) was accomplished the induced pluripotent stem cell fate. In a preliminary study, we have fol- 20 years ago by the mere modulation of the culture conditions. Recently, lowed thirty-four likely targets of the ESCC miRNAs during reprogramming. we converted adult germline stem cells (GSCs) into germline-derived A large number of these targets are expressed and suppressed by the ESCC pluripotent stem (gPS) cells. GSCs are unipotent cells of the testis that are miRNAs during reprogramming. These targets can be categorized into spe- capable of not only self-renewing, but also giving rise to sperm. Like ES cific cellular processes including cell cycle, epithelial to mesenchymal transi- cells, GSCs exhibit significant levels of Oct4 and Klf4, but low endogenous tion (EMT), vesicular transport, and epigenetic regulation, providing insights expression Sox2 and c-Myc. To better understand the reprogramming pro- into how these processes may be regulated during reprogramming. Further cess, we sought to identify factors that mediate reprogramming at higher evaluation of the EMT targets confirmed a role for the ESCC miRNAs in efficiency. We established an assay based on Oct4 reactivation to screen promoting a mesenchyamal to epithelial transition during reprogramming. nuclear fractions from extracts of pluripotent cells. BAF chromatin remodel- We are currently performing larger screens for miRNA-mRNA pairs that ing complexes containing the Brg1 protein have been shown to be not only promote the induction of pluripotency as well as other cell fate transitions. essential for early embryonic development, but also, as we have previously An update of these ongoing experiments will be presented. shown, paramount in enhancing the efficiency of reprogramming somatic cells to pluripotency mediated by the quartet. As knockdown of Brg1 leads to differentiation of ES cells, we investigated the early effect of Brg1 knockdown by assessing the impact of RNA interference on the expression levels of key pluripotency factors. We show that Brg1 knockdown leads

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Plenary III – Tissue Stem Cell Origins WNT SIGNALLING AND THE REGULATION OF MUSCLE STEM CELL FUNCTION Supported by Lieber Institute for Brain Development Rudnicki, Michael A. Thursday, June 16, 9:00 – 11:30 a.m. Ottawa Hospital Research Institute, Ottawa, ON, Canada SPECIFICATION AND MAINTENANCE OF Satellite cells in adult skeletal muscle are a heterogeneous population NEURAL CREST CELL FATE composed of stem cells and committed progenitors. We undertook a gene expression analysis of satellite stem cells towards identifying signaling Bronner, Marianne pathways that regulate their function. Our experiments identified the non- Dept. of Biology, California Institute of Technology, Pasadena, CA, USA canonical Wnt-receptor Fzd7 as being specifically expressed in quiescent satellite stem cells. Wnt7a was examined as a candidate receptor for Fzd7 The neural crest is a population of multipotent, migratory progenitor cells because of its expression and role during embryonic and adult myogen- that forms at the border of neural and non-neural ectoderm in vertebrate esis. We found that Wnt7a was markedly upregulated in newly formed embryos. These cells then migrate from the neural tube along defined myofibers during regenerative myogenesis, and that the Fzd7 receptor is pathways, populate numerous sites and differentiate into diverse cell types necessary for Wnt7a binding at the surface of myogenic cells. Recombinant — including melanocytes, sensory and autonomic neurons. We propose Wnt7a protein dramatically stimulated the symmetric expansion of satellite that a putative gene regulatory network of interacting transcriptional stem cells and that this expansion required Fzd7 and Vangl2, both compo- regulators and downstream effector genes confers properties like multipo- nents of the planar cell polarity (PCP) signaling pathway. Moreover, Wnt7a tency and migratory capacity to nascent neural crest cells. We are testing induced polarized localization of Vangl2 at opposite poles in pairs of divid- this hypothesis by systematically perturbing a subset of the transcription ing cells, in a manner consistent with Wnt7a activating PCP signaling. Over factors involved in early neural crest specification and examining the effect expression of Wnt7a during muscle regeneration resulted in an impressive of these perturbations on likely downstream genes in order to test the pre- enhancement of the regeneration process, generating more fibers of bigger dicted interrelationships. By isolating cis-regulatory regions of genes in this caliber, independent of an effect on myoblast proliferation or differentia- putative neural crest regulatory network to identify neural crest enhancers, tion. Importantly, Wnt7a over-expression resulted in a large expansion of we have identified additional inputs to the network and determined which the satellite stem cell population, and Wnt7a deficiency resulted in impaired interactions are direct. The results suggest that a series of gene regulatory maintenance of the satellite cell compartment. Therefore, Wnt7a signaling circuits are involved in inducing the migratory neural crest cell population, through the planar cell polarity pathway controls the homeostatic level of maintaining its stem cell properties for a time and finally leading to progres- satellite stem cells and hence regulates the regenerative potential of muscle. sive differentiation. In differentiated myofibers, we observed that Wnt7a binding to Fzd7 di- rectly activates the Akt/mTOR growth pathway thereby inducing myofibre hypertrophy. Notably, the Fzd7 receptor complex was associated with DEVELOPMENTAL PROCESS OF PI3kinase specifically in differentiated myotubes and myofibres, but not in HEMATOPOIETIC STEM CELL EXPLAINED myoblasts, and was required for Wnt7a to activate the Akt/mTOR growth pathway. Wnt7a/Fzd7 activation of this pathway was completely indepen- Nishikawa, Shin-Ichi dent of IGF-receptor activation. Together, these experiments demonstrate RIKEN Center for Developmental Biology, Kobe, Japan that Wnt7a/Fzd7 activates distinct pathways at different developmental stages during myogenic lineage progression, and together identify a novel Hematopoietic stem cell (HSC) is the most extensively studied stem cell, but non-canonical anabolic signaling pathway for Wnt7a and its receptor Fzd7 yet its developmental pathway in has not been fully explained. in skeletal muscle. In this ISSCR, I will present our current understanding about the defini- tive HSC (dHSC) differentiation course. At present, 3 distinct stages were identified between nascent mesoderm and dHSC. The primitive mesoderm exofoliated from the primitive streak express both PDGFRα and Flk1. The ENDOTHELIAL ORIGINS OF HEMATOPOIETIC first event for the lineage divergence to dHSC is expression of a transcrip- STEM CELLS tion factor (TF), Etv2/ER71 in this primitive mesoderm. As null mutation of 1 2 3 etv2/er71 gene results in complete block of differentiation of EC and HPC Dzierzak, Elaine , Yokomizo, Tomomasa , Robin, Catherine , lineages, all dHSC are derived from Etv2+ cells. Etv2 loss results in complete Speck, Nancy4 differentiation arrest of the primitive mesoderm to Flk1-single positive 1Erasmus Medical Center Erasmus Stem Cell Institute, Rotterdam, lateral mesoderm. Thus induced Flk1+ lateral mesoderm is fated to EC, as Netherlands, 2Cancer Science Institute of Singapore, Singapore, Singapore, Etv2 directly induce molecular network that is involved in maintenance the 3Erasmus Stem Cell Institute, Rotterdam, Netherlands, 4University of EC fate. The second step of divergence of hematopoietic lineages occurs by Pennsylvania School of Medicine, Philadelphia, PA, USA additional expression of Runx1. When lateral mesoderm expresses Runx1, Hematopoietic stem cells (HSC) are the source of all blood cells in the adult it enters the Runx1+ second stage that acquires the potential to give rise and are the basis of cell replacement therapies for blood-related genetic to dHSC. Which genes are regulated by Runx1 at this stage is yet to be diseases and leukaemias. Since HSC numbers are limited and do not readily investigated. Runx1+ cells are then incorporated into caudal-ventral region expand, an understanding of the developmental processes that lead to of embryo together with other angioblasts fated to EC to form vascular their generation in the embryo can provide insight into possible ex vivo system. This migration occurs before the circulation begins. Probably, only manipulation of HSCs. The first HSCs are generated in the aorta-gonad-me- a small proportion of the progeny of Runx1+ cells are integrated into the sonephros (AGM) region at midgestation in the mouse embryo and at week vascular system, particularly at dorsal aorta and omphalloallantoic ves- 4-6 in human gestation. AGM HSCs are generated following the anatomical sels. This is the third stage that is designated as hemogenic EC. An array appearance of hematopoietic clusters associated with the lumenal wall of of recent video-imaging studies including ours clearly showed that CD45+ the aorta and vitelline umbilical arteries. The relationship of HSCs to these cells bud out from the hemogenic EC both in vitro and in vivo. We are fully clusters and the identification of the direct precursor cell to HSCs is an area convinced that this is the scheme to start with. of intense research focus. The direct precursor(s) to HSCs is suggested to be the endothelial or mesenchymal cells directly underlying the hematopoietic clusters, or hematopoietic cells from the circulation that lodge in the clusters

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and undergo a maturational event. To begin to understand how clusters are formed we have developed a whole mount immunostaining method by Concurrent Session I Track A – which we can temporally map and quantitate all hematopoietic clusters in normal (and hematopoietic defective) mouse embryos that have been made Human iPS and Embryonic Stem Cells transparent. We have localized HSC activity to the hematopoietic clusters. Supported by Development Genetic data show that HSC generation requires the Runx1 transcription factor in VE-cadherin (endothelial) cells. Deletion of Runx1 in these cells Thursday, June 16, 1:30 – 3:15 p.m. also leads to an absence of vascular hematopoietic clusters, supporting the notion of hemogenic endothelial cells as the precursors to HSCs/clusters. SOX2, STEM CELLS AND CELLULAR Visual proof that HSCs arise from aortic endothelium comes from time lapse REPROGRAMMING vital imaging data of the AGM. Remarkably, HSCs arise directly from endo- thelial cells of the dorsal aorta in a natural transdifferentiation event. These Hochedlinger, Konrad data and data on the molecules in the surrounding AGM microenvironment Harvard University and Massachusetts General Hospital, Boston, MA, USA involved in the induction and growth of AGM HSCs will be discussed. I will present data on the transcriptional and epigenetic comparison of embryonic stem cells and induced pluripotent stem cells in mouse, which revealed remarkable similarities but also subtle differences that affect LGR5 STEM CELLS IN SELF-RENEWAL AND developmental potential. I will also talk about our recent analysis of Sox2 CANCER expression patterns in adult tissues. We have identified Sox2-positive cells in multiple adult epithelial tissues where it has not previously been described. Clevers, Hans C. I will provide evidence to show that these Sox2+ cells are adult stem cells Hubrecht Institute, Royal Netherlands Academy of Arts and Sciences & that are essential for adult tissue turnover and survival. University Medical Centre Utrecht, Utrecht, Netherlands The intestinal epithelium is the most rapidly self-renewing tissue in adult mammals. We originally defined Lgr5 as a Wnt target gene, transcribed ESTABLISHMENT OF CFTR-EXPRESSING in colon cancer cells. Two knock-in alleles revealed exclusive expression EPITHELIAL FROM PLURIPOTENT STEM CELLS of Lgr5 in cycling, columnar cells at the crypt base. Using an inducible Cre knock-in allele and the Rosa26-LacZ reporter strain, lineage tracing Wong, Amy P.1, Pasceri, Peter1, Thompson, Tadeo O.2, Ratjen, experiments were performed in adult mice. The Lgr5+ve crypt base columnar Felix3, Gonska, Tanja4, Bear, Christine4, Ellis, James1*, Rossant, cells (CBC) generated all epithelial lineages throughout life, implying that Janet1* it represents the stem cell of the small intestine and colon. Similar obser- 1Development & Stem Cell Biology, Hospital for Sick Children, Toronto, vations were made in hair follicles and stomach epithelium. Single sorted ON, Canada, 2Ontario Human Induced Pluripotent Stem Cell Facility, +ve Lgr5 stem cells can initiate ever-expanding crypt-villus organoids in 3D Hospital for Sick Children, Toronto, ON, Canada, 3Physiology & +ve culture. Tracing experiments indicate that the Lgr5 stem cell hierarchy is Experimental Medicine, Hospital for Sick Children, Toronto, ON, Canada, maintained in these organoids. We conclude that intestinal crypt-villus units 4Molecular Structure and Function, Hospital for Sick Children, Toronto, are self-organizing structures, which can be built from a single stem cell ON, Canada in the absence of a non-epithelial cellular niche. The same technology has now been developed for the Lgr5+ve stomach stem cells. *Equal contributions Intestinal cancer is initiated by Wnt pathway-activating mutations in genes Cystic fibrosis (CF) is a fatal disease that affects airway ciliated epithelial such as APC. As in most cancers, the cell of origin has remained elusive. cells caused by mutations in the cystic fibrosis transmembrane conductance Deletion of APC in stem cells, but not in other crypt cells results in progres- regulator (CFTR) gene. Limited availability of CF epithelium remains sively growing neoplasia, identifying the stem cell as the cell-of-origin of a major roadblock for the potential development of therapeutic drugs adenomas. Moreover, a stem cell/progenitor cell hierarchy is maintained in a preclinical setting. Induced pluripotent stem (iPS) cells hold great in early stem cell-derived adenomas, lending support to the “cancer stem promise for patient-specific disease modeling, drug discovery and cell”-concept. Fate mapping of individual crypt stem cells using a multicolor regenerative medicine. At present, there are no efficient and reliable Cre-reporter revealed that, as a population, Lgr5 stem cells persist life-long, methods to generate airway epithelia from human pluripotent cells. This yet crypts drift toward clonality within a period of 1-6 months. Lgr5 cell project aims to develop an efficient step-wise differentiation protocol divisions occur symmetrically. The cellular dynamics are consistent with a for generating functional airway epithelia from patient CF-iPS cells. To model in which the resident stem cells double their numbers each day and date, iPS lines from 3 patients homozygous for the F508 CF mutation stochastically adopt stem or TA fates after cell division. Lgr5 stem cells are have been generated and extensively characterized. Efficient generation interspersed between terminally differentiated Paneth cells that are known of endoderm progenitors (>75%) from embryonic stem (ES) cells has to produce bactericidal products. We find that Paneth cells are CD24+ and been established using Activin-A. Lung epithelial progenitors have been express EGF, TGF-a, Wnt3 and the Notch ligand Dll4, all essential signals induced with high concentration of FGF2 and carefully timed treatment of for stem-cell maintenance in culture. Co-culturing of sorted stem cells epithelial morphogens with up-regulation of airway epithelial genes highest with Paneth cells dramatically improves organoid formation. This Paneth following low stimulation of Nodal or FGF18. Following manipulation of cell requirement can be substituted by a pulse of exogenous Wnt. Genetic cell microenvironment by air liquid interface to mimic maturation of the removal of Paneth cells in vivo results in the concomitant loss of Lgr5 stem developing lung, the cells form dome-like structures (a hallmark of epithelial cells. In colon crypts, CD24+ cells residing between Lgr5 stem cells may transport properties). Immunofluorescent staining of the cells show tight represent the Paneth cell equivalents. We conclude that Lgr5 stem cells epithelial junctions formed (ZO1+) with apical localization of CFTR. compete for essential niche signals provided by a specialized daughter cell, Electrophysiological assessments using a modified SPQ assay and Ussing the Paneth cell. chamber analysis are underway to assess functional establishment of ES cell-derived CFTR-expressing epithelia. These patient-specific epithelial cells may provide a renewable source to identify novel therapeutics.

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THE UBIQUITIN PROTEASOME SYSTEM lesser requirement for the Sox2. We tested whether inactivation of Nanog, Oct4 and Sox2 triggers specific differentiation programs. Downregulation of REGULATES SELF RENEWAL AND Nanog in several hESC lines led to the induction of neuroectodermal mark- DIFFERENTIATION OF MOUSE EMBRYONIC ers, while markers of other germ layers were undetectable. This signature STEM CELLS was further enhanced following stimulation with Noggin, while incuba- tion with bFGF or serum led to the emergence of neuronal cell types. Two Buckley, Shannon M., Strikoudis, Alexandros, Aifantis, Iannis distinct patterns were observed following the Oct4 inactivation. Extraem- Dept. of Pathology, Howard Hughes Medical Institute/NYU School of bryonic lineages - trophectoderm and endoderm - were induced in H1 and Medicine, New York, NY, USA several other hESCs, while the neuroectodermal lineage was induced in H9 hESCs. Activation of Bmp4 signaling was both necessary and sufficient for Pluripotent embryonic stem (ES) cells, as well as tissue-specific adult stem the induction of extraembryonic lineages, and blocking the endogenous cells, have the unique ability to both self-renew and differentiate. Tran- signaling redirected differentiation toward the neuroectodermal fates. scriptional regulation has been in the center of stem cell self-renewal vs. Downregulation of Sox2 did not result in activation of lineage-specific differentiation conundrum as transcriptional networks in ES cells are well differentiation. While both RNA and protein levels of Sox2 were decreased substantiated. However, accumulating evidence suggests that regulation more than tenfold, the levels of Nanog and Oct4 remained unchanged. of gene transcription is not the whole answer to the question of stem cell Stable Sox2-deficient clones, capable of giving rise to teratomas in vivo, fate decisions. Our laboratory has strong evidence pointing at post-trans- were derived and maintained in culture for more than 20 passages. At the lational modifications, and more specifically regulation by the ubiquitin- same time, an increase in differentiated cells was also observed, indicating proteasome system (UPS) as a molecular mechanism controlling stem cell that hESCs with reduced levels of Sox2 are prone to spontaneous differen- differentiation. Indeed, we have shown that the E3 ubiquitin ligase Fbw7 tiation. Marker analysis combined with testing in directed differentiation plays an integral role in adult stem cell self-renewal, and preliminary data assays revealed a bias toward the endomesodermal fates. To understand demonstrates that Fbw7 expression is dynamically regulated during ES cell how the global molecular networks of hESCs are regulated by Nanog, Oct4 differentiation. Additionally, RNAi-mediated Fbw7 silencing inhibits ES cell and Sox2, we analysed transcriptome dynamics following gene inactiva- differentiation, while maintaining pluripotency and self-renewal of ES cells. tion using microarrays. Clustering of 1,717 genes perturbed by at least one Inhibition of the proteasome with MG132 treatment leads to apoptosis, but shRNA, revealed that only a small fraction of genome requires all three more importantly differentiation with a loss of core transcription regula- regulators for the maintenance of the proper expression. Large cluster of tors including Nanog, suggesting an important role of the proteasome in genes was co-regulated by Nanog and Oct4, and a large Oct4-specific suppressing differentiation. To investigate the role of the UPS in ES cells, we cluster was also detected. The Nanog-specific cluster was relatively small have performed a siRNA-based ubiquitin screen targeting 636 members of and only six Sox2-specific genes were identified, suggesting that the role of the ubiquitin family including E3 ligases and their antagonist deubiquitinat- Sox2 is to reinforce the Nanog and Oct4-mediated transcriptional networks. ing enzymes to elucidate the role of the UPS in ES cell self-renewal and In summary, we show that Nanog and Oct4 are essential for hESC self- differentiation. Using a Nanog-GFP reporter mouse ES cell line, we have renewal. Inactivation of these genes in hESCs triggers lineage-specific pro- identified 20 genes, which decrease Nanog expression in self-renewing grams different from those activated in mESCs. Sox2, while not absolutely conditions and 17 genes that inhibit differentiation of ES cells when treated required, does suppress spontaneous differentiation by reinforcing the with retinoic acid. In the screen we identified Psmd14, a deubiquitinase Nanog and Oct4-mediated regulatory networks. Our results suggest that and also a subunit of the proteasome lid. Knockdown of Psmd14 results manipulating levels of self-renewal regulators such as Nanog and Sox2, is in differentiation and increased apoptosis. In order to further elucidate the a useful way to facilitate differentiation of hESCs into clinically relevant cell substrates and pathways altered by the UPS members, we used proteomic type. approaches. Utilizing tandem affinity purification methods coupled with mass spectrometry analysis for UPS members identified in the screen, we revealed Psmd14 interacting partners, including a number of subunits of the proteasome confirming the role of UPS in self-renewal. Our results suggest REPROGRAMMING UNDER OPTIMIZED the UPS, which can modulate protein function, plays a key role in regulat- CONDITIONS REVEALS A SHORTENED ROUTE ing embryonic stem cell fate decisions. Further characterization of identified TO PLURIPOTENCY ligases and their substrates may further unravel mechanisms of pluripoten- cy, self-renewal, and differentiation, as well as, provide clues of mechanisms Pei, Duanqing disrupted in cancers. South China Institute for Stem Cell Biology and Regenerative Medicine, Guangzhou Institutes of Biomedicine and Health, Chinese Academy of Sciences, Guangzhou, China NANOG-OCT4-SOX2 REGULATORY MODULE IN Reprogramming of somatic cells to a pluripotent state by defined factors HUMAN EMBRYONIC STEM CELLS represents a major breakthrough in life sciences as cell fate can be studied rationally and systematically. However, the mechanism and the technol- Wang, Zheng, Oron, Efrat, Nelson, Brynna, Ivanova, Natalia ogy of reprogramming remains poorly understood and inefficient, thus at Department of Genetics, Yale University, New Haven, CT, USA its infancy. We hypothesized that somatic cell reprogramming by defined Understanding pluripotency and differentiation in ESCs is critical to our factors is a collaborative process between the defined factors and the cell ability to produce specific cell types for medicine and research. Studies in culture condition. We have shown recently that vitamin C when added to the mouse identified transcription factors Nanog, Oct4 and Sox2 as the the medium and inhibition of TGFb signaling can greatly enhance repro- master regulators of pluripotency. However, it remains unclear whether and gramming, underscoring the significance of the extracellular environment how these genes contribute to the regulation of hESCs. We utilized lentiviral in reprogramming. In this presentation, we present evidence that further shRNAs to inactivate Nanog, Oct4 and Sox2 in hESCs. To determine the formulation of reprogramming specific media can lead to shortened routes general requirement of each gene, the shRNA-transduced RFP+ hESCs were towards pluripotency. Under optimized conditions, mouse fibroblasts can co-cultured with GFP+ control hESCs and the RFP+/GFP+ cell ratios were be reprogrammed to a pluripotent state with ultra-high efficiency (~10% monitored over time. Rapid decreases in RFP+/GFP+ ratios were observed at day 8) with Oct4/Sox2/Klf4, or at lower efficiency with Oct4 only. Mo- for all Nanog and Oct4 shRNAs, indicating that these genes are essential, lecular studies reveal an interesting interplay between chromatin remodeling while a slower kinetics was observed for the Sox2 shRNAs, suggesting a factors, cell cycle regulators and signaling mediators that reduce the barrier of reprogramming to achieve pluripotency with less time and less factors.

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Detailed Program and Abstracts – Thursday, June 16

This new paradigm not only provides a rationale to further improve iPSC ENGINEERING THE 3D MICROENVIRONMENT generation but also simplifies the conceptual understanding of reprogram- ming by defined factors. OF MULTI-CELLULAR PLURIPOTENT STEM CELL AGGREGATES FOR DIRECTED DIFFERENTIATION AND MORPHOGENESIS Concurrent Session I Track B – Bratt-Leal, Andres M., Kinney, Melissa A., Carpenedo, Richard L., Sargent, Carolyn Y., Hammersmith, Katy A., Kepple, Kirsten L., Stem Cells & Tissue Engineering McDevitt, Todd C. Supported by Ontario Stem Cell Initiative (OSCI) Biomedical Engineering, Georgia Tech / Emory University, Atlanta, GA, USA Thursday, June 16, 1:30 – 3:15 p.m. Stem cell differentiation is sensitive to various global and local environmen- TOWARDS AN INTEGRATED SUSPENSION- tal cues that direct cell fate decisions to divergent phenotypes. Pluripotent stem cells (i.e. ESCs & iPSCs), inherently capable of differentiating into cell BASED PLURIPOTENT STEM CELL INDUCTION, types comprising all three germ lineages (ecto-, endo-, and mesoderm), EXPANSION AND DIRECTED DIFFERENTIATION recapitulate many aspects of early development and serve as a robust cell source for the development of cell-based diagnostics and regenerative PLATFORM medicine therapies. However, the efficiency of directing pluripotent stem Zandstra, Peter W., Ungrin, Mark, Fluri, David, Clarke, Geoff, cell differentiation to specific phenotypes, particularly in 3D aggregates Baptista, Ricardo commonly referred to as “embryoid bodies” (EBs), is often limited by the inability to effectively control the presentation of morphogenic cues. Thus, Donnelly Center, University of Toronto; McEwen Centre for Regenerative in order to better understand and ultimately control ESC differentiation, we Medicine, University Health Network, Toronto, ON, Canada have focused on systematically controlling global and local parameters of EB Producing the anticipated cellular mass needed for many human plu- microenvironments and examining the effects on the assembly, intercellular ripotent stem cell (hPSC)-associated applications remains challenging. communication and morphogenesis of multi-cellular 3D stem cell aggre- Primarily empirical approaches for culture scale-up must be replaced with gates. Embryoid bodies (EBs) were formed by rotary orbital suspension cul- rational and quantitative bioprocess design strategies employing cell- and ture directly (2x10^5 cells/mL; 10 mL/100 mm dish) or initially aggregated molecular-level analysis of rate-limiting steps in scalable cell expansion and in PDMS microwells (100-1000 cells/well; Aggrewell™) for ~24 hours prior directed differentiation technologies. We have defined a novel bioprocess to being introduced and maintained for up to 14 days of differentiation in optimization parameter (L; targeted yield Loss) and, with carboxyfluo- rotary orbital suspension. Rotary orbital speed was varied between 20-65 rescein succinimidyl ester (CFSE) cell division tracking and functional and rpm for different cultures. Microparticles (~1-20 µm diameter; MPs) of dif- phenotypic fate monitoring of hPSC expansion and differentiation, used it ferent materials (i.e. gelatin, PLGA, agarose) were introduced by forced ag- to identify and overcome key suspension bioprocess bottlenecks. First, we gregation in microwells with different ratios of MPs to cells at the time of EB demonstrated that aggregate size control (using arrays of microwells) could formation. Gene expression was assayed by PCR arrays and qRT-PCR, while enable adherent-equivalent growth rates of hPSC in feeder-free matrix- protein expression was assessed by immunoblotting and/or immunostain- free defined suspension culture. Optimal aggregate size (30-200 cells per ing followed by microscopy and cytometry. Hydrodynamic forces, imparted aggregate) could be predicted across different hESC and induced hPSC lines by rotary orbital suspension culture, affected not only the formation and based on line-specific input parameters such as population growth rates and morphology of ESC spheroids, but also influenced global transcriptional single cell survival frequencies. Taking advantage of the observation that activity and subsequent differentiation of the cell populations, particularly cell aggregation phenomena change during -mediated repro- to endo- and mesoderm lineages. The modulation of ESC differentiation ap- gramming, we have overcome the need to serially dissociate aggregates peared to be mediated at least in part by temporal changes in intracellular during pluripotent cell expansion by integrating fibroblast reprogramming beta-catenin signaling between the different groups examined. In addi- into our suspension bioprocess. Exogenously regulated transgene activation tion, microparticles of varying size and different materials were physically enabled high density (>5x106 cell/mL) cell production processes wherein entrapped within aggregates of ESCs in a dose-dependent manner without a majority of the cells could be continuously generated as single cells or adversely affecting cell viability and could serve as delivery vehicles for as loosely adherent aggregates. Kinetic genome-wide analysis of suspen- morphogens, such as small molecules and growth factors. Microparticle- sion vs. adherent culture reprogramming identified significant differences mediated delivery of differentiation factors, including retinoic acid, BMP4 in adhesion factor gene expression (e.g., cadhereins, extracellular matrix- and VEGF, induced different gross morphological and phenotypic effects on related genes) between the two conditions; these and other culture specific ESC differentiation compared to soluble delivery methods and moreover, changes resulted in a marked acceleration in the rate and efficiency of the presence of different types of materials alone appeared to modulate EB reprogrammed cell generation in suspension culture. Applying these tech- differentiation. Altogether, these examples demonstrate that combining dif- nologies to directed differentiation protocols allowed us to demonstrate that ferent levels of macro- (i.e. hydrodynamic) and micro- (i.e. particles) control stable pluripotent cell populations (such as hESC) can, via predominantly to regulate stem cell 3D environments can be used to more effectively instructive mechanisms, undergo ~5 proliferation divisions to generate direct differentiation and morphogenesis. It is expected that the develop- definitive endoderm capable of further development along pancreatic and ment of multi-scale techniques to direct pluripotent stem cell differentiation hepatic trajectories. In contrast, when suspension bioreactor generated iPSC will benefit the biomanufacturing of stem cell derivatives for regenerative were directly transitioned into differentiation induction protocols, target cellular therapies and in vitro cell based diagnostic technologies, as well as cell generation was concomitant with, at least initially, a significant culture enable engineering of tissues directly from stem cells. condition-dependent cell selection process. Imposing technologies with increased control over temporal and spatial heterogeneity has revealed fundamental insights into the growth and differentiation of PSC, moving us closer to strategies capable of efficiently and robustly generating PSC and their derivatives for drug screening and therapy.

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Detailed Program and Abstracts – Thursday, June 16

DIRECTED INDUCTION OF CHONDROGENIC understanding of tubulogenesis in modular environments as well as a clini- cally relevant alternative to natural materials for regenerative medicine. In CELLS FROM MURINE DERMAL FIBLOBLAST this study, we utilized tunable hyaluronic acid (HA) hydrogels to investigate CULTURE WITHOUT GOING THROUGH A ECFCs morphogenesis. First, we demonstrate that ECFCs express CD44 PLURIPOTENT STEM CELL STATE and CD168, specific receptors for HA and produce hyaluronidase (Hyal) in response to vascular endothelial growth factor (VEGF). We then show that Otani, Hidetatsu1, Hiramatsu, Kunihiko1, Yoshikawa, Hideki2, VEGF and matrix stiffness co-regulate ECFCs morphogenesis. High level of Tumaki, Noriyuki1 VEGF is required to initiate vascular morphogenesis and to activate mem- 1Department of Bone and Cartilage Biology, Osaka University Graduate brane type -1 matrix metalloproteinase (MT1-MMP), MMP-1, and MMP-2, School of Medicine, Suita, Japan, 2Department of Orthopaedic Surgery, to enable ECFCs migration. With decrease in substrate elasticity, which Osaka University Graduate School of Medicine, Suita, Japan corresponds to a loose ultrastructure, we observe decreased MMPs expres- sion and increased cellular elongation, with intracellular vacuole extension It has been reported that the combined transduction of two reprogram- and coalescence to open lumen compartments. RNAi studies confirmed that ming factors (c-Myc and Klf4) and one chondrogenic factor (SOX9) induces MT1-MMP is required to enable ECFCs migration and that ECFCs sense chondrogenic cells directly from mouse dermal fibroblast (MDF) culture matrix stiffness through a signaling cascade leading to the activation of (J Clin Invest, doi: 10.1172/JCI44605). The purpose of this study is to Cdc42. Next, we show that vacuole and lumen formation are RGD dose- determine whether the cells in the MDF culture go through pluripotent dependent and are recognized by the ECFCs through integrin α5β1 and αVβ3 stem cell state during induction into chondrogeic cells by transduction of subunits. Integration of MMP-sensitive-peptide to the HA hydrogels en- c-Myc, Klf4 and SOX9. For real-time monitoring of the pluripotent stem abled ECFCs to sprout, branch, and form complex vascular networks. Along cell state, we used the mice bearing the BAC transgene in which EGFP is the culture period, increased expression of hyaluronidase isoforms Hyal-1, inserted into the 5’-untranslated region of the Nanog gene (Nanog-GFP) -2, as well as MMPs by the vascular networks result in decreased matrix (Nature, 2007;448:313). MDFs were prepared from Nanog-GFP mice at stiffness and release of the encapsulated growth factors, demonstrating the 6-week old, transduced with c-Myc, Klf4 and SOX9 using retroviral vectors, modularity of the synthetic environment. Finally, to determine the function- replated into 6-well plates, and subjected to time-lapse GFP observation for ality of the engineered vascular networks, matured and stabilized vascular 14 days. Each whole well was scanned with 8 x 8 images every 8 hours. As constructs were subcutaneously implanted into nude mice. By two weeks previously reported, nodules composed of cells with polygonal-morphology post implantation, 85% of the HA hydrogels were degraded and replaced appeared by 7 days after transduction. Nodules were stained with alcian by macrophages and tissue ingrowth. Blood vessels in various sizes are blue intensely at 14 days after transduction, suggesting that chondro- found at the periphery and center of the hydrogels. Most microvasculatures genic cells were induced. We found around 15 chondrogenic nodules per at the periphery of the gels are of murine origin. Two types of microvessels each well, whereas we detected no GFP fluorescence by the time-lapse at the center of the hydrogels are found: about 60% of the blood vessels observation during this period. As for the positive control, we transduced contain both human ECFCs and host cells, while the remaining vessels con- the Nanog-GFP MDFs with c-Myc, Klf4, Oct3/4, and Sox2, and detected tain only human ECFCs. Perfusion with blood cells is detected in both types GFP fluorescence 3 weeks after transduction. Next, we collected RNAs from of microvasculature, demonstrating that the implanted ECFCs participate MDF culture for every other day for 10 days after transduction of c-Myc, in the angiogenesis of the host vasculature and form functional human Klf4, and SOX9, and subjected to RT-PCR expression analysis. Expression vascular networks that anastomose with the host vasculature to form levels of Col1a1 and Col1a2 (fibroblast markers) decreased and expression functional vessels (vasculogenesis). Collectively, we show that the signaling level of Col2a1 (chondrocyte marker) increased in MDFs transduced with pathways of ECFCs vascular morphogenesis can be precisely regulated in a c-Myc, Klf4, and SOX9, compared with those in MDFs without transduc- synthetic matrix, resulting in a robust and functional microvasculature useful tion. Expression levels of Nanog were markedly low in MDFs transduced for the study of stem cell vascular biology and towards a range of vascular with c-Myc, Klf4, and SOX9, compared with that in ES cells. These results disorders and approaches in tissue regeneration. collectively suggest that chondrogenic cells were induced directly from MDF culture by transduction of c-Myc, Klf4, and SOX9 without going through a pluripotent stem cell state. ENGINEERED LUNGS FROM BIO-SCAFFOLDS AND PULMONARY CELLS CONTROLLING MORPHOGENESIS OF Niklason, Laura E. ENDOTHELIAL PROGENITORS TO GENERATE Yale University, New Haven, CT, USA FUNCTIONAL MICROVASCULATURE IN A Adult lung tissue is limited in its ability to regenerate, meaning that severely SYNTHETIC MATRIX damaged lungs can only be treated by lung transplantation, a procedure that is hampered by organ rejection and infection. In order to regenerate Hanjaya-Putra, Donny1, Bose, Vivek1, Shen, Yu-I1, Yee, Jane1, lung tissues in vitro, adult rat lungs are carefully decellularized to remove Khetan, Sudhir2, Burdick, Jason A.2, Gerecht, Sharon1 cellular components, while preserving the extracellular matrix and retain- 1Department of Chemical and Biomolecular Engineering, Johns Hopkins ing the hierarchical branching structures of airways and vasculature. A Physical Science Oncology Center, Johns Hopkins University, Baltimore, novel lung culture bioreactor is utilized to culture pulmonary epithelium MD, USA, 2Department of Bioengineering, University of Pennsylvania, and vascular endothelium on acellular lung matrices and produce tissue Philadelphia, PA, USA engineered lungs. Seeded and cultured epithelium displays remarkable hierarchical organization within the lung matrix, while seeded endothelial The generation of functional vascular networks by endothelial colony- cells efficiently repopulate the vascular compartment. In addition, mechani- forming cells (ECFCs) has the potential to improve treatment for vascular cal characteristics of engineered lungs are similar to those of native tissues. diseases and to facilitate successful transplantation of tissue-engineered Engineered lungs are implantable in vivo, and can participate efficiently in organs. ECFCs are recruited from a bone marrow niche to the site of gas exchange, demonstrating that this approach is effective for regenerat- vascularization, where cues from the extracellular matrix (ECM) instigate ing functional lung tissue in vitro. vascular morphogenesis. Although this process has been elucidated using natural matrix, little is known about vascular morphogenesis of ECFCs in synthetic matrices where properties can be tuned, towards both the basic

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Detailed Program and Abstracts – Thursday, June 16

Concurrent Session I Track C – Neural Stem Cells COMPARATIVE FUNCTIONAL GENOMIC RNAI SCREEN OF EMBRYONIC AND NEOPLASTIC Supported by StemCells, Inc. NEURAL STEM CELLS Thursday, June 16, 1:30 – 3:15 p.m. Aviv, Tzvi1, Ward, Ryan1, Clarke, Ian1, Ketela, Troy2, Moffat, ENVIRONMENTAL FACTORS REGULATING Jason2, Dirks, Peter1 NEURAL STEM CELL LINEAGE PROGRESSION 1Developmental & Stem Cell Biology Program, Hospital for Sick Children Toronto, Toronto, ON, Canada, 2Molecular Genetics, University of Toronto, Kokovay, Erzsi, Wang, Jenny, Goderie, Susan K., Shen, Qin, Toronto, ON, Canada Temple, Sally The median survival of glioblastoma (GBM) patients is under two years. Neural Stem Cell Institute, Rensselaer, NY, USA Regeneration of GBMs following surgery and chemotherapy may be at- Neural stem cells (NSCs) produce transit amplifying stem cells and these tributed to a rare population of self-renewing cells termed cancer stem in turn generate neuronal and glial progeny. Environmental signals, such cells (CSCs). Recently, we demonstrated that CSCs from human and as growth factors and cytokines, can activate lineage progression. In prior mouse GBMs share similar gene expression pattern with non-tumorigenic studies we showed that NSCs express the CXCR4 receptor and respond neural stem cells (NSCs) of embryonic origin. However, normal NSCs are to the cytokine SDF1 produced by vascular endothelial cells, using this to non-tumorigenic when injected into immunocompromised mice whereas home to the vascular niche that stimulates proliferation. Hence, there are CSCs generate a tumor that phenocopies the original tumor. To elucidate fundamental parallels between the NSC system and the hematopoietic stem molecular mechanisms that drive self-renewal of embryonic and tumori- cell (HSC) system, which uses SDF1/CXCR4 to regulate stem cell homing genic stem-cells we employed genome-wide short-hairpin RNAi (shRNAi) into the bone marrow niche. More recently we have extended that parallel knockdown to identify genes required for proliferation of NSCs and CSCs. to show that another HSC-homing associated factor, VCAM1, is impor- We infected p53 deficient NSCs or CSCs derived from p53 deficient mice tant for the maintenance of NSCs and the normal niche cytoarchitecture. GBMs with lentiviral shRNAi library of 80,000 sequences. We assessed Anti-VCAM1 antibody delivered using osmotic minipumps in vivo into the the distribution of shRNAi cassettes that integrated into the genomes of forebrain causes dramatic disruption of the subventricular zone stem cell recipient cells immediately after infection (day 0) using custom designed niche and interrupts neuroblast migration to the olfactory bulb. Knockdown microarrays. Nine and fourteen days later, we sampled genomic DNA from of VCAM-1 causes loss of stem cell renewal markers and stimulates their infected cells and compared the distribution of shRNAi cassettes to that differentiation into neurons. In contrast, increasing VCAM1 levels sustains of day 0. Our analyses revealed 557 essential genes for which the shRNAi the NSC state. In addition to their role in homeostasis, both these cytokine cassettes ‘dropped out’ (DO) during passages of all NSC lines including systems are important for inflammatory responses, providing a mechanism lines of embryonic and neoplastic origin. This list of genes includes expected for NSC responsiveness to disease or damage. targets, such as components of the DNA, RNA and protein synthesis ma- chineries. Importantly, 80 kinases are also significantly enriched among DO genes including many brain-specific kinases, emphasizing the complex and tight signaling regulation on NSC proliferation. Enrichment of kinase targets SYNAPTIC DEFECTS IN HUMAN NEURONS was observed also among 202 shRNAi cassettes that dropped out in GBM DERIVED FROM AUTISM SPECTRUM lines exclusively. These kinases include cancer related genes such as Chek1 DISORDERS PATIENTS and Lkb1 and GBM related Ephrin receptors. Currently, we are validating the anti-proliferative effects of shRNAi that target essential kinases in all Muotri, Alysson R. NSC lines and in CSC exclusively. Validated hits will be further examined to Pediatrics/Cellular & Molecular Medicine, UCSD, La Jolla, CA, USA determine their influence on cell-cycle status and differentiation potential of the cells. Our work identifies many genes with potential regulatory rules Autism spectrum disorders (ASD) are complex neurodevelopmental diseases in self-renewal and differentiation of NSCs. Genes required for proliferation in which different combinations of genetic mutations may contribute to the of neoplastic CSCs may reveal the molecular mechanisms at play during phenotype. Using Rett syndrome (RTT) as an ASD genetic model, we devel- tumorigenesis and relapse and may provide new targets for selective inhibi- oped a culture system using induced pluripotent stem cells (iPSCs) from RTT tion of CSCc in brain tumors. patients’ fibroblasts. RTT patients’ iPSCs are able to undergo X-inactivation and generate functional neurons. Neurons derived from RTT-iPSCs had fewer synapses, reduced spine density, smaller soma size, altered calcium signaling and electrophysiological defects when compared to controls. RTT PEERING INTO STEM CELLS IN LIVE BRAIN: neurons were also used to test the effects of drugs in rescuing synaptic A NEW SUBTYPE OF NEUROGENIC defects. Finally, RTT-iPSC differentiated into neural precursor cells supported PROGENITOR IN MOUSE NEOCORTEX an increase amount of somatic mutations caused by de novo L1 retrotrans- poson insertions. To identify common cellular and molecular pathways Wang, Xiaoqun, Kriegstein, Arnold involved in autism, we derived neurons from other ASD-iPSCs. Interestingly, University of California San Francisco, San Francisco, CA, USA similarly to RTT-neurons, we also observed defects in a subset of ASD neu- rons. These data suggest that perturbation in common molecular pathways A hallmark of mammalian brain evolution is cortical expansion, which can lead to neuronal alterations involved in the etiology of ASDs. The over- reflects an increase in the number of cortical neurons. This is established lap phenotypes observed in the iPSC-derived human neurons between ASD by the type of progenitor cells present in embryonic brain and by their patients provide evidence of an unexplored developmental window, before number of neurogenic divisions. In developing rodent neocortex, radial glias disease onset, where potential therapies could be successfully employed. generate intermediate progenitor cells that migrate to the subventricular Our model recapitulates early stages of a human neurodevelopmental dis- zone where they divide symmetrically to produce neurons. Recent studies ease and represents a promising cellular tool for drug screening, diagnosis revealed a new class of radial glia-like cells in human brain, oRG cells, which and personalized treatment. reside in the outer subventricular zone and are progenitors. Expansion of the subventricular zone and the appearance of oRG cells may have been es- sential evolutionary steps leading from lissencephalic to gyrencephalic neo- cortex. Here we show that progenitor cells resembling oRG cells are present

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Detailed Program and Abstracts – Thursday, June 16 in mouse embryonic neocortex, and arise from asymmetric divisions of A NOVEL APPROACH TO REGENERATIVE radial glia. Time-lapse imaging reveals that the cells undergo self-renewing asymmetric divisions to generate neurons. Moreover, the mouse oRG cells MEDICINE: REVERSION OF THE POSTMITOTIC undergo mitotic somal translocation, whereby the centrosome moves into STATE BY TRANSIENT DUAL KNOCKDOWN OF a varicosity in the basal process during interphase, and the nucleus follows TUMOR SUPPRESSORS prior to mitosis. Our finding of oRG cells in rodent brain fills a gap in our understanding of the cellular basis for neocortical expansion. Corbel, Stephane1, Pajcini, Kosta1, Mourkioti, Foteini1, Sage, Julien2, Blau, Helen M.1 1Microbiology and Immunology, Stanford Univ, Stanford, CA, USA, A TRANSCRIPTIONAL MECHANISM LINKING 2Pediatrics and Genetics, Stanford Univ, Stanford, CA, USA SOX2 AND PARAHOX GENES IN ADULT Many human tissues, are postmitotic, like heart and pancreas, and are TISSUES characterized by an absence of stem cells and reduced regenerative capac- ity. In contrast, newts and zebrafish possess extremely robust regenerative McKay, Ronald D.1, Kuzmichev, Andrei2, Kim, Suel-Kee2 potential, as strikingly demonstrated by their ability to re-grow their hearts 1Lieber Institute, Baltimore, MD, USA, 2NINDS, NIH, Bethesda, MD, USA and entire limbs or fins. Recent lineage tracing has demonstrated that this is not due to the specialization of pluripotent cells at the site of amputation, Sox2 is a central regulator of stem cells in pluripotent cells and in the adult but instead the regeneration process entails dedifferentiation of dedicated brain. Here we use conditional expression to show that Sox2 induction rap- cartilage, bone and muscle cells to a proliferative state. We reasoned that idly increased the number of stem cells in the intestine. Before cell prolifera- the reactivation of such naturally occurring reparative pathways in humans tion occurred, Sox21 was induced and the parahox gene Cdx2 repressed. could constitute an advantageous regenerative mechanism, as the cells that Our results suggest that inhibition of Cdx2 is a general mechanism of Sox2 proliferate would maintain their specialized identity. We recently used an action in pluripotent and colon cells. When Oct4 was induced in the adult evolutionary approach to elucidate differences between humans and newts intestine, cell proliferation occurred but there was no change in the number that might be fruitfully exploited in tissue regeneration. Newts inactivate of stem cells or expression of either Sox21 or Cdx2. Specific genetic target- the cell cycle regulator Rb by phosphorylation, and lack the regulator ing showed that Sox2 acts in a cell autonomous manner in intestinal stem p19/Arf encoded by the Ink4a locus (which first arose in chickens). We cells. The extraordinary conservation of the parahox gene cluster suggests hypothesized that transient inactivation of these two tumor suppressors that the developmental regulation of these master genes is a specific role of by introducing siRNAs into postmitotic fully differentiated mononuclear Sox2 that should be further explored in stem cells. mammalian muscle cells, known as myocytes, could induce the cells to dedifferentiate. Our data support this hypothesis and show that inactiva- tion of p19/Arf and Rb in skeletal muscle, leads to cell cycle re-entry (BrdU labeling), loss of differentiation properties, upregulation of the cytokinetic Concurrent Session I Track D – machinery, and cell division. Using live single cell laser micro-dissection- catapulting, we obtained definitive evidence that si-treated postmitotic Stem Cells and Regeneration myocytes that express myogenin-GFP (the point of no return when re-entry Supported by The Hospital for Sick Children, Research Institute into the cell cycle cannot be induced by growth factors) when isolated and plated in culture, they divided. Transient suppression of both p19/Arf Thursday, June 16, 1:30 – 3:15 p.m. and Rb by siRNAs induced proliferation, whereas neither alone sufficed. Myoblast colonies arose that retained the ability to differentiate and fuse DERMAL STEM CELLS: FROM THE SKIN TO into myofibers and regenerate damaged muscles upon in vivo transplanta- THE SPINAL CORD tion. Our data show that differentiation of mammalian cells can be reversed by temporary inactivation of tumor suppressors, p19/Arf and Rb, and the Miller, Freda resulting dedifferentiated cells are capable of contributing to skeletal muscle Hospital for Sick Children, University of Toronto, Toronto, ON, Canada repair in vivo. This approach of inducing specialized cells of known identity We previously identified a multipotent neural crest-related stem cell from to dedifferentiate - i.e., to take just one step back and divide - could prove in rodent and human skin that we called SKPs for skin-derived precursors. useful in generating cells of diverse tissue types, both in the intact tissue situ These cells are able to generate mesenchymal cells types such as dermal and in tissue culture. The cells generated by temporarily removing the fibroblasts, adipocytes and chondrocytes, and peripheral neural cells such brakes on the cell cycle may serve as a potent adjunct/alternative to iPS in as Schwann cells. We recently defined the cells-of-origin for SKPs and modeling human diseases and for screening for ameliorative drugs, as they show that they are sox2-positive dermal hair follicle precursors that can may exhibit a more mature phenotype. reconstitute the dermis and induce hair morphogenesis. This talk will focus upon the endogenous biology of these cells, including molecular pathways that regulate their self-renewal and maintenance, and on the possibility DISEASE MODELING WITH INDUCED that these cells can generate cell types that are never seen in the dermis. PLURIPOTENT STEM CELLS REVEALS THE Moreover, the use of transplanted SKP-derive Schwann cells for spinal cord regeneration will also be presented. PATHOGENESIS OF SOMATIC MOSAICISM IN AN NLRP3-DRIVEN AUTOINFLAMMATORY SYNDROME Tanaka, Takayuki, Saito, Megumu, Takahashi, Kazutoshi, Yamanaka, Shinya, Nakahata, Tatsutoshi Kyoto University, Kyoto, Japan Background: Derivation of induced pluripotent stem cells (iPSC) from fibro- blasts enables us to study diseases under a new prospect. So far, disease modeling in neural or cardiac disorder has been reported, but there has

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Detailed Program and Abstracts – Thursday, June 16

been no report regarding immunologic disorder. Chronic Infantile Neuro- APPLYING REGENERATIVE CONCEPTS FOR logic Cutaneous and Articular (CINCA) syndrome is a severe autoinflamma- tory disease caused by gain-of-function mutations of NLRP3 gene. Mutant THE CNS FROM THE SALAMANDER TO NLPR3 protein in macrophages causes overproduction of IL-1 beta, which MAMMALIAN NEURAL STEM CELLS

leads to systemic inflammation. We previously identified patients of this 1 2 2 3 syndrome with somatic mosaicism of NLRP3 mutation. It has been contro- Zhu, Yu , Meinhardt, Andrea , Ader, Marius , Niesche, Marco , 2 1 versial (1) whether only blood cells with NLRP3 mutation have abnormal Stec, Agnieszka , Tanaka, Elly function of cytokine secretion or all the cells in mosaic patients carry anoth- 1Max Planck Institute of Molecular Cell Biology and Genetics, DFG er unknown mutation and abnormal function and (2) whether the NLRP3- Research Center for Regenerative Therapies, Technical University of mutant macrophages with relatively small ratio actually cause the systemic Dresden, Dresden, Germany, 2DFG Research Center for Regenerative inflammation. Results: We obtained fibroblasts from two male patients of Therapies, Technical University of Dresden, Dresden, Germany, CINCA syndrome with somatic mosaicism. Fibroblasts were transduced 3Department of Neurosurgery, University Clinic, Technical University with lentiviral vector encoding ecotropic receptor Slc7a1 and with retroviral Dresden, Dresden, Germany vector encoding OCT4, SOX2, KLF4 and cMYC. Compact embryonic stem Our lab has traditionally studied how the salamander can undertake such (ES)-cell like colonies emerged after 2 weeks and we obtained more than comprehensive, functional regeneration of the central nervous system. 20 iPSC lines. We obtained both wild-type and mutant clones from each From these studies we have found that regeneration occurs at least in patient’s fibroblasts. After evaluating transgene silencing by quantitative part through a multipotent neural stem cell. At the onset of regeneration, RT-PCR, we selected 3 mutant and wild type iPSC clones respectively for ependymo-glial cells in the spinal cord revert to form a simple neuroepithe- further analysis. All the clones expressed human ES-cell markers. iPSCs are lium that grows as a tube and replays the early events of neural develop- differentiated into CD14+ macrophages via CD34+KDR+ hematopoietic ment. The formation of this self-organizing tube appears to be a critical progenitors. Mutant iPSC-derived macrophages secreted higher amount of part of the regeneration process. Based on this knowledge we have started IL-1 beta when stimulated with lipopolysaccharides or infected with Listeria to model these regenerative properties in human and mouse neural stem monocytogenes, while wild-type clones did not. Moreover, when mutant cells by focusing on producing self-organizing neuroepithelial tubes in three and wild type macrophages are mixed, they produced significantly more IL- dimensional culture. We have found that culturing mouse and human ES 1beta than only mutant cells did, suggesting the interaction of mutant and cells in classical epithelial conditions promotes efficient differentiation to wild type macrophages in a patient’s body. neuroepithelia. In the human system our conditions promote eye field formation. Further culture of these human neuroepithelia in activin results in quantitative differentiation of cells into retinal pigment epithelia whereas REGULATION OF MOUSE EPIDERMAL culture in FGF promotes neural retina formation. In the mouse system we STEM CELLS NICHE HETEROGENEITY BY A have defined the conditions for generating spinal cord neuroepithelia with different dorsal-ventral identities. These studies show that culture conditions MOLECULAR CLOCK promoting epithelial integrity are an important contributor to efficiently Aznar Benitah, Salvador direct cells along the neural lineage. This work was supported by grants from the International Foundation for Paraplegia, the DFG Collabora- Differentiation and Cancer, Center for Genomic Research (CRG), ICREA tive Research Center 655: From Cells to Tissues, the Max Planck Institute Researcher, Barcelona, Spain for Molecular Cell Biology and Genetics and the Center for Regenerative Epidermal stem cells ensure that skin homeostasis is maintained. In murine Therapies, Dresden. skin, epidermal stem cells cluster at specific niches where, at steady state conditions, they undergo cycles of dormancy and activation. Upon the requirement of cellular replenishment, epidermal stem cells egress the niche and proliferate for a limited number of times to subsequently feed into the Concurrent Session I Track E – Stem Cell Signaling differentiated compartment. However, at each round of morphogenesis, only a subset of stem cells becomes active, suggesting that heterogeneous Thursday, June 16, 1:30 – 3:15 p.m. stem cell responsive states coexist within the same niche. Using a circa- dian clock fluorescent reporter mouse model, we show that the dormant HAIR FOLLICLE STEM CELL PROVIDE A epidermal stem cell niche contains two populations of stem cells at opposite FUNCTIONAL NICHE FOR MELANOCYTE STEM phases of the clock, which are differentially predisposed to respond to niche CELLS homeostatic cues. In dormant niches, the core molecular clock circuitry transcriptionally modulates the expression of stem cell regulatory genes to Nishimura, Emi create two coexisting stem cell populations, one predisposed, and one less Dept of Stem Cell Biology, Medical Research Institute, Tokyo Medical and prone, to activation. Unbalancing this equilibrium of epidermal stem cells, Dental University, Chiyoda-ku, Japan through conditional epidermal deletion of a core clock component, results in a long-term progressive accumulation of non-responsive stem cells, pre- In most stem cell systems, the organization of the stem cell niche and the mature impairment of tissue self-renewal, and a significant reduction in the anchoring matrix required for stem cell maintenance are largely unknown. development of squamous cell carcinomas. We propose that the molecular Melanocyte stem cells (MSC) and hair follicle stem cells (HFSC), which are clock machinery finetunes the spatiotemporal behavior of epidermal stem originally derived from a completely different developmental origin, are cells within their niche, and that perturbation of this mechanism affects tis- located in the bulge area of mammalian hair follicles. Our previous studies sue homeostasis and the predisposition to neoplastic transformation. indicated that the niche plays dominant role in MSC fate determination, while the underlying mechanisms and the correlation with HFSCs remain unclear. We report here that collagen XVII (COL17A1/BP180/BPAG2), a hemidesmosomal transmembrane collagen, is highly expressed in hair follicle stem cells (HFSCs) and is required for the maintenance not only of HFSCs but also of melanocyte stem cells (MSCs), which do not express Col17a1 but directly adhere to HFSCs. Mice lacking Col17a1 show prema- ture hair graying and hair loss. Analysis of Col17a1 null mice revealed that COL17A1 is critical for the self-renewal of HFSCs through maintaining their

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Detailed Program and Abstracts – Thursday, June 16 quiescence and immaturity, potentially explaining the mechanism underly- TAILORED SYNTHETIC SURFACES TO CONTROL ing hair loss in human COL17A1 deficiency. Interestingly, Col17a1 null mice show defective TGF-β production by HFSCs and targeted TGF-b type HUMAN PLURIPOTENT STEM CELL FATE II receptor (Tgfbr2) deficiency in the melanocyte lineage causes incomplete Klim, Joseph R.1, Li, Lingyin2, Wrighton, Paul J.3, Kiessling, Laura maintenance of melanocyte stem cell immaturity and results in premature L.4 hair graying. These data demonstrate that the TGF-b signaling pathway is 1Cell and Molecular Biology, University of Wisconsin Madison, Madison, the critical niche factor that regulates melanocyte stem cell immaturity and WI, USA, 2Systems Biology, Harvard Medical School, Boston, MA, USA, quiescence. Finally, forced expression of COL17A1 in basal , 3Biochemistry, University of Wisconsin Madison, Madison, WI, USA, including HFSCs, in Col17a1 null mice rescues MSCs from premature differ- 4Chemistry, University of Wisconsin Madison, Madison, WI, USA entiation and restores TGF-β signaling, demonstrating that HFSCs function as a critical regulatory component of the MSC niche. Human pluripotent stem cells hold great promise for the fields of regenera- tive medicine, developmental biology, and drug discovery. To exploit the full potential of these cells, we must elucidate the molecular mechanisms HIGH CONTENT, SINGLE CELL ANALYSIS OF underlying their proliferation and differentiation. Therefore, defined culture conditions are needed. While defined media capable of maintaining human EMBRYONIC STEM CELL DIFFERENTIATION BY pluripotent stem cells are known, identifying chemically-defined, robust MASS CYTOMETRY substrata has proven more difficult. To this end, we recently employed

1 2 3 surface arrays to screen substrates that engage a wide variety of different Zunder, Eli R. , Linderman, Michael D. , Lujan, Ernesto , cell surface receptors. From these arrays, we identified surfaces that support 1 1 1 Simonds, Erin F. , Bendall, Sean C. , Bodenmiller, Bernd , the long-term propagation of human pluripotent stem cells. The effective Behbehani, Greg K.1, Qiu, Peng4, Plevritis, Sylvia K.4, Wernig, substrates displayed heparin-binding peptides, which can interact with cell Marius3, Nolan, Garry P.1 surface glycosaminoglycans. Conversely, pluripotent stem cells cultured on 1Baxter Laboratory in Stem Cell Biology, Department of Microbiology and surfaces displaying only integrin-binding peptides were unable to maintain Immunology, Stanford University, Stanford, CA, USA, 2Computer Systems an undifferentiated population of cells. To exploit this phenomenon, we Laboratory, Stanford University, Stanford, CA, USA, 3Institute for Stem Cell have patterned synthetic surfaces with discrete areas presenting heparin- Biology and Regenerative Medicine, Stanford University, Stanford, CA, binding peptides or integrin-binding peptides. These tailored surfaces USA, 4Department of Radiology, Stanford University, Stanford, CA, USA provide spatial control over cell fate decisions. Our tailored surfaces also provide insights into the molecular pathways regulating the actin-myosin Differentiation of pluripotent stem cells into specific cell types is essential for contractility associated the poor cloning efficiency of human embryonic therapeutic applications, but cellular heterogeneity in differentiating cultures stem cells. The loss of cell-cell interactions is known to lead to actin-myosin complicates analysis and the identification of underlying molecular mecha- contractility in human embryonic stem cells. Our results demonstrate that nisms. To investigate differentiation at the single-cell level, mouse embry- cell-extracellular matrix interactions also affect actin-myosin contractility onic stem (mES) cells were analyzed by mass cytometry after inducing dif- in human embryonic stem cells. Specifically, we propose that simultane- ferentiation by leukemia inhibitory factory (LIF) withdrawal and retinoic acid ous engagement of the glycosoaminoglycan and integrins is required to (RA) treatment. Mass cytometry is a novel flow cytometry technique that enhance integrin signaling and prevent actin-myosin contraction. Since this uses rare earth metal isotopes instead of fluorophores for antibody labeling requirement is specific to undifferentiated cell state, it can be exploited in and subsequent epitope detection, increasing the number of parameters combination with our tailored surfaces to separate differentiated cells from available relative to fluorescence-based flow cytometry, and eliminating pluripotent cells. We anticipate that these findings will yield advances in overlap between measurement channels. Over a five-day time course of culturing human pluripotent cells and insight into the signaling mechanisms mES cell differentiation, 34 markers of pluripotency, differentiation, cell underlying pluripotency and differentiation. cycle, and cellular signaling were monitored simultaneously at the single-cell level. To analyze the high-dimensional mass cytometric data, Spanning-tree Progression Analysis of Density-normalized Events (SPADE) was used, an unsupervised approach that identifies unique cell populations in n-dimen- THE CANCER STEM CELL MARKER CD133 IS sional space and arranges them hierarchically into a minimum spanning tree REGULATED BY HISTONE DEACETYLASE 6 (Peng et al., Phenotypically determined self-organization of flow cytometry AND FUNCTIONS TO STABILIZE BETA-CATENIN data with spanning-tree progression analysis of density-normalized events, submitted). The resulting SPADE trees reveal potential pathways for lineage IN WNT SIGNALING FOR SUPPRESSION OF progression over the time course of mES cell differentiation, from multiple CANCER CELL DIFFERENTIATION “mES-like” populations, to intermediately-differentiated populations, to 1 2 1 SPADE tree branches representing each of the three germ layers. The cel- Mak, Anthony B. , Stewart, Jocelyn M. , Kittanakom, Saranya , lular state of each population in the progression tree, revealed by signaling Chen, Ginny I.1, Curak, Jasna1, Gingras, Anne-Claude1, Mazitschek, molecules and cell cycle status, identifies key points for small molecule Ralph3, Stagljar, Igor1, Neel, Benjamin G.2, Moffat, Jason1 intervention that can direct differentiation or maintain pluripotency. Mass 1Molecular Genetics, University of Toronto, Toronto, ON, Canada, cytometry coupled with SPADE analysis is a powerful new approach to the 2Medical Biophysics, University of Toronto, Toronto, ON, Canada, 3Center study of pluripotency, differentiation, and cellular signaling, and it has great for Systems Biology, Chemical Biology Platform, Massachusetts General potential for additional systems such as induced pluripotent stem (iPS) cell Hospital, Boston, MA, USA reprogramming, transdifferentiation, and directed differentiation. The CD133 epitope, AC133, has been a marker associated with stem cells and cancer stem cells (CSCs) from various tissue types. CD133 positive cancer cells are more resistant to traditional cancer treatments, including both chemo- and radio-therapy, representing a subpopulation of cells for cancer relapse. In addition, CD133 expression marks a poor prognosis in certain cancer types. Despite its utility as a primitive cell marker, regula- tion of CD133 expression is poorly understood and there is little evidence for a functional role of CD133. We wanted to identify protein interactions of CD133 to guilt its function by association. We used the mammalian

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Detailed Program and Abstracts – Thursday, June 16

affinity purification and lentiviral expression (MAPLE) system (Mak et al, Mol Cell Proteomics, 2010) coupled with mass spectrometry analyses and Plenary IV – identified the microtubule-associated histone deacetylase 6 (HDAC6) as a direct physical interaction partner of CD133. Targeting HDAC6 using RNAi Stem Cells and Cancer – Biology and Drug Development knockdown or the HDAC6-specific small molecule inhibitor tubacin resulted Thursday, June 16, 4:00 – 6:00 p.m. in CD133 protein downregulation. We further demonstrate that HDAC6 negatively regulates trafficking of CD133 into endosomal compartments for PHENOTYPIC PLASTICITY IN HUMAN SOMATIC subsequent lysosomal degradation. Targeting CD133 directly or indirectly via HDAC6 using RNAi decreases proliferation in vitro and tumor xenograft CELLS growth in vivo by triggering cell differentiation in colon and ovarian cancer Tlsty, Thea D., Roy, Soma, Gascard, Philippe, Dumont, Nancy cells. This change in cell state is due to our finding that CD133 promotes Dept. of Pathology, University of California, San Francisco, San Francisco, the stabilization and nuclear localization of beta-catenin, which positively CA, USA regulates Wnt signaling. Our study suggests that CD133 and HDAC6 function in regulating Wnt signaling and are involved in suppressing cancer The ability to bypass negative growth signals and participate in wound cell differentiation. Given that CD133 is a CSC marker in a wide range of healing is a fundamental function of somatic stem cells. Such cells have cancers, targeting the CD133/HDAC6/Wnt axis may provide a means to long been isolated from adult tissue and exhibit a restricted, tissue-specific treat multiple cancer types. potency for generating multiple cell fates. For example, hematopoietic stem cells reconstitute the immune system in radiated individuals but are not used to generate skin grafts. Recent reports have characterized special- CONTROL OF STEM CELLS BY DIET AND ized fetal populations (i.e. amniotic cells) with a broader potency as well as mesenchymal populations with various potentials to express multiple cell SYSTEMIC FACTORS IN THE DROSOPHILA fates. It is currently unknown if any cell within the adult body escapes the OVARY potency restrictions which are imposed as an organism undergoes develop- ment and differentiation. This topic will be discussed. Drummond-Barbosa, Daniela, Ables, Elizabeth, Hsu, Hwei-Jan, LaFever, Leesa Department of Biochemistry and Molecular Biology, Johns Hopkins ENDOCYTOSIS STEM CELLS AND CANCER Bloomberg School of Public Health, Baltimore, MD, USA di Fiore, Pier Paolo Despite intense research on how stem cells are regulated by intrinsic factors and local signals from their niches, much less is understood about how stem Istituto FIRC di Oncologia Molecolare and Istituto Europeo di Oncologia, cells sense and respond to hormones, diet or other environment factors. Milan, Italy Our pioneering work showed that ovarian stem cells adjust their prolifera- Signaling receptors are internalized following their engagement by cognate tion rates in response to diet in Drosophila, and that insulin signals are key ligands, in a process that has been traditionally considered crucial for mediators of this process. Specifically, brain-derived insulin-like peptides long-term attenuation of receptor signaling. However, recent evidence directly modulate the division of germline stem cells (GSCs) via PI3K/FOXO has pointed also to an important role of endocytosis in conferring spatial at the level of G2 and independently of the niche. More recently, we found and temporal dimensions to signaling (the signaling endosome concept). that insulin-like peptides also regulate GSC maintenance/numbers via a dis- It is thus predicted that subversion of the endocytic machinery might play tinct mechanism. Insulin-like peptides control niche size by modulating the an important role in cancer. Of particular interest, in this contention, is an ability of cap cells within the niche to respond to Notch ligands. In addition, endocytic protein called Numb. Numb is a cell fate determinant that by insulin signals affect the levels of E-cadherin at the cap cell-GSC junction in asymmetrically partitioning at mitosis controls binary cell fate decisions. In a Notch-independent manner. We have also obtained several pieces of evi- human breast and lung cancers, there is frequent loss of Numb expression. dence indicating that insulin-like peptides are not the only systemic signals This causes alterations in two major downstream pathways. On the one required for the stem cell response to diet. In fact, we have recently found hand, lack of Numb allows for unchecked signaling activity of the Notch that the steroid hormone ecdysone plays a key role in GSC regulation. Our receptor. On the other, lack of Numb causes attenuation of the p53 signal- results show that ecdysone, which is produced in later developing follicles, ing pathway. Tumors cells displaying loss-of-Numb expression are addicted directly regulates adult GSC proliferation and self-renewal independently to this event and to its molecular consequences. Our recent results point of insulin signaling. Ecdysone controls GSCs through a functional interac- to the mammary stem cell (MSC) compartment as the cellular “target” of tion with the chromatin remodeling factors ISWI, an intrinsic epigenetic Numb misregulation in breast tumors. We have developed a technology to factor required for GSC fate and activity, and Nurf301, the largest subunit cultivate ad purify mammary stem cells. In normal MSC, Numb is asymmet- of the ISWI-containing NURF chromatin remodeling complex. Further, we rically partitioned at mitosis. This in turn dictates the replicative fate, in that found that the direct action of ecdysone on GSCs is crucial for their normal the Numb(+) cell remains quiescent (and retains MSC capabilities), whereas response to bone morphogenetic protein (BMP) signals from the niche. the Numb(-) cell acquires a progenitor fate and undergoes rapid symmet- We propose the model that ecdysone produced by more differentiated ric divisions. The control of Numb over MSC fate is executed through the ovarian cells under favorable conditions acts on GSCs in part to enhance ability of Numb of silencing Notch signaling and maintaining high levels their response to niche signals as a positive feedback mechanism. Our of p53 in the MSC. Lack of Numb in tumor MSC causes a switch form the findings demonstrate the complexity of effects of systemic factors on stem asymmetric to the symmetric mode of division, thus forcing both daughter cells, including modulation of niche size and function as well as interac- cells to assume the same replicative fate. Our understanding of how Numb tions with intrinsic stem cell factors. Diet and other environmental factors is mechanistically involved in all these aspects will be discussed. drive evolution of all living organisms. Given the crucial roles of stem cells in the development, maintenance and/or function of all tissues, the ability of these cells to sense and respond to physiological changes elicited by diet and other factors is likely universally conserved.

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Detailed Program and Abstracts – Thursday, June 16 and Friday, June 17

CLINICAL IMPLICATIONS OF EPIGENETIC we detected. Collectively, our data illustrate that leukemic progression can occur in either a linear or branching fashion, with multiple genetic subclones REGULATORS OF SELF RENEWAL evolving either in succession or independently, respectively. The discovery Clarke, Michael F. that specific genetic events influence L-IC frequency and that genetically distinct L-IC evolve through a complex evolutionary process indicates that Stanford Institute for Stem Cell Biology and Regenerative Medicine, Palo a close connection must exist between genetic and functional heterogene- Alto, CA, USA ity. These findings indicate that there may be more commonalities between Stem cells are defined by the ability to divide and give rise to a new stem clonal evolution and CSC models of cancer than previously thought. Finally, cell (self-renewal), as well as the ability to give rise to the differentiated our study points to the need to develop effective therapies to eradicate all cells of an organ, and thus are the only long-lived cell population in many genetic subclones in order to prevent further evolution and recurrence. tissues. The polycomb protein Bmi1 has been shown to be essential for the self renewal of normal stem cells in multiple organs including the blood, brain and breast. We have identified modifiers of Bmi1 which are dif- ferentially expressed between hematopoietic stem cells (HSCs), which are Plenary V – Reprogramming and Fate Conversion endowed with self-renewal potential, and multipotent progenitors, which cannot self renew. These modifiers counteract the action of the Polycomb Supported by The New York Stem Cell Foundation (NYSCF) factor. Overexpression of these modifiers alone markedly impairs stem cell Friday, June 17, 9:00 – 11:30 a.m. self renewal. One of these modifiers is expressed in a specific region of Chromosome 21 frequently associated with Downs Syndrome (DS), and INDUCTION OF PLURIPOTENCY BY DEFINED our data suggests that in DS mouse model that contains an extra copy of this modifier-containing region of chromosome 21, stem cell compart- FACTORS ments of the blood and breast are reduced. Epithelial tumors consist of a Yamanaka, Shinya heterogeneous population of cancer cells that differ in their apparent state Center for iPS Cell Research and Application (CiRA), Kyoto University, of differentiation, suggesting that solid tumors might represent aberrant Kyoto, Japan organs containing a cancer stem cell population that maintains the ability to self-renew. Indeed, using a xenograft model of human breast, colon and Induced pluripotent stem (iPS) cells were originally generated from mouse head and neck cancers, a phenotypically-distinct subset of the cancer cells and human fibroblasts by the retroviral introduction of Oct3/4, Sox2, c- (cancer stem cells) has been found to have the exclusive ability to form Myc, and Klf4. iPS cells are similar to embryonic stem (ES) cells in morphol- tumors. The remaining cancer cells, which often form the bulk of the tumor, ogy, proliferation, gene expression, and most importantly, pluripotency. are unable to self-renew or sustain tumorigenesis. The Bmi1-modifiers are Patient-specific iPS cells provide us with unprecedented opportunities in also differentially expressed by cancer stem cells compared to the other can- disease research, drug screening, toxicology, and cell transplantation thera- cer cells. Enforced expression of the Bmi1 modifiers inhibits tumorigenesis. pies. Banks of iPS cells from individuals with HLA homozygous alleles may also provide another unprecedented source for regenerative medicine. iPS cells have many variations in comparison to ES cells. In addition to THE GENETIC DIVERSITY OF LEUKEMIA fibroblasts, iPS cells can be generated from various somatic cells, such as hepatic cells, gastric epithelial cells, keratinocytes, neural cells, peripheral INITIATING CELLS OCCURS THROUGH A blood cells, and cord blood cells. In addition to retroviral transduction, iPS COMPLEX EVOLUTIONARY PROCESS cells can be integration-freely generated by adenoviruses, sendaiviruses, plasmids, transposons, recombinant proteins and synthetic mRNA. Several 1 1 2 Dick, John E. , Notta, Faiyaz , Mullighan, Charles chemicals and inhibitors have been shown to enhance iPS cell generation. 1Princess Margaret Hospital, University Health Network, and Dept Some of the original four factors can either be omitted or replaced by other of Molecular Genetics, University of Toronto, Toronto, ON, Canada, genes or chemicals. Finally, each induction experiment can result in 100 or 2Department of Pathology, St. Jude Children’s Research Hospital, Memphis, more independent iPS cell clones. TN, USA These various iPS cells may vary significantly in regard to each cell’s peculiar The cellular and molecular basis for the heterogeneity that exists within characteristics, such as the efficacy of in vitro directed differentiation and the individual cells that make up a tumour is not well understood. Genetic a propensity to produce tumors. In fact, the origins of iPS cells have a diversity combined with clonal evolution is proposed to drive much of the profound effect on their tumorigenicity. It is therefore necessary to deter- functional heterogeneity within tumours. Often proposed as a mutually mine the best origins, the best induction protocols, and the best methods to exclusive alternate model, the cancer stem cell (CSC) model postulates that evaluate iPS cell clones and subclones for each application of iPS cells. Our heterogeneity arises because of epigenetic differences in gene expression recent study show that L-Myc can take c-Myc’s place and generate safer that result in the tumour being organized as a cellular hierarchy sustained iPS cells at higher efficiencies. Here, significance of epigenome analyses, by a CSC at the apex. As a first step to determine if these two concepts of including methylome, histone modifications, and imprinting increases as the cancer could be harmonized, we direct addressed whether human leukemia integration-free method is becoming practical.It is also important to note was composed of genetically diverse subclones and whether functionally that an iPS cell clone can be heterogeneous despite the fact that all the cells defined leukemic initiating cells (L-IC) are genetically diverse within an within the clone are derived from a single progenitor cell. This is because the individual sample. We focused on Ph+ ALL because it is considered a single process requires multiple cell divisions which cannot be completed by the four clinical entity where genome-wide DNA copy number alteration (CNA) exogenous factors alone. Additional factors including p53 and Rb pathways profiling was recently undertaken. A robust Ph+ B-ALL xenograft system are considered to play various roles in achieving full reprogramming. Obtain- was developed that enabled combined genetic and functional studies of the ing a better understanding of the reprogramming mechanisms will therefore genetic diversity of functionally defined L-IC derived from diagnostic pa- facilitate the generation of more uniform iPS cells in the future. tient samples. We found that genetic diversity occurs in functionally defined L-IC and that many diagnostic patient samples contain multiple genetically distinct L-IC subclones that are related through a complex evolutionary process. Reconstruction of the genetic events that were present in the sub- clones combined with assay of the functional properties of each subclone in xenografts enabled us to reconstruct the genetic ancestry of the subclones

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Detailed Program and Abstracts – Friday, June 17

THE DIRECT REPROGRAMMING OF SOMATIC myeloid genes in pre-B cells induced to transdifferentiate by C/EBPa. In addition, the expression of the human orthologues of the Tet2 target genes CELL NUCLEI TO AN EGG OR OOCYTE PATTERN correlated with the level of Tet2 expression in acute myeloid leukemias OF GENE EXPRESSION (AML), and Tet2high samples were more differentiated than Tet2low samples, and also relatively hypomethylated. In addition, we found that C/EBPa Gurdon, John induces the upregulation of Tet2 during transdifferentiation and that Tet2 University of Cambridge, Wellcome Trust/Cancer Research UK Gurdon in turn induces an increase of hydroxy-methylated cytosines at target gene Institute, Cambridge, United Kingdom promoters. Furthermore our data indicate that C/EBPa and Tet2 proteins As cells differentiate they become increasingly committed to their final associate, suggesting that C/EBPa recruits Tet2 to its target genes, a finding specialization. However, the stability of their differentiated state can be which could explain the specificity of the Tet2 effects. Our data suggest experimentally reversed by procedures that include nuclear transfer, cell that during blood cell differentiation Tet2 endows cells with the ability to fusion, and iPS technology. This talk will discuss chromatin changes associ- rapidly respond to cell fate instructive signals through the erasure of repres- ated with nuclear transfer to eggs and oocytes. When trying to analyze the sive epigenetic memory mediated by DNA methylation. mechanisms responsible for nuclear reprogramming and gene activation, it is helpful to have an experimental system in which the reprogramming takes place rapidly and efficiently. If the efficiency of the nuclear repro- TRANSPOSON-MEDIATED REPROGRAMMING gramming is judged by the ability of nuclei of cells from one specialized PROVIDES A POWERFUL TOOL FOR path of differentiation to generate cells of another unrelated pathway, then the efficiency of reprogramming by eggs in second meiotic metaphase is UNDERSTANDING SOMATIC CELL about 30%. However, the events that immediately follow the transplanta- REPROGRAMMING TO PLURIPOTENCY tion of somatic nuclei to eggs are difficult to analyze because, immediately after nuclear transfer to eggs, the major activity of a transplanted nucleus is Nagy, Andras to replicate DNA, and transcription starts only after many hours (mammals) Mount Sinai Hospital, Samuel Lunenfeld Research Institute, Toronto, ON, or many cell divisions (amphibia). When somatic nuclei are transplanted to Canada the germinal vesicle of an oocyte in first meiotic prophase, a very efficient Somatic cell reprogramming with a few defined transcription factors to activation of previously quiescent pluripotency genes takes place. This pluripotency is a several weeks long process. The driving forces behind this experimental system is being used to identify the components of oocytes phenomenon and the cascade of events are very poorly understood. It is that induce pluripotency gene reactivation and which elicit major changes however crucial to uncover the fine details of this process in order to com- to the structure and composition of chromatin. An important and necessary prehend the true property of these induced pluripotent stem cells (iPSCs) part of the reprogramming process is the substitution of the somatic linker and so better tailor their future therapeutic and disease study use. histone H1o by the oocyte or embryo-specific linker histone B4 H1foo. Two years ago we developed a reprogramming method utilizing a transpo- This is a necessary change to the chromatin without which gene reactiva- son-mediated delivery of the reprogramming . Beyond the ability tion does not take place. When chromatin has become decondensed by of seamless removal of the transgenes once pluripotent stem cells have action of linker histone exchange and other substitutions, the genome of been generated, this system has additional unique features. For example transplanted nuclei appears to be able to respond to basic transcriptional when it combined with the doxycycline inducible transgene expression sys- factors and not to require gene-specific transcription factors. The combina- tem we found that these transgenes are very efficiently regulatable by add- tion of structural chromatin changes and an insensitivity to gene-specific ing or withdrawing doxycycline. In vivo differentiated somatic cells derived transcription factors begins to approach a mechanistic explanation for the from iPSCs can be reprogrammed to “secondary” iPSCs (2ºiPSc) by simply reprogramming of transplanted nuclei by first meiotic prophase oocytes. adding doxycycline to the culture medium. Somatic cell lines produced with The eventual understanding of reprogramming by oocytes will be to this method frequently return to 2ºiPSc in a “population” manner, which provide a time course of the key events which take place when a somatic allows us to study the cascade of molecular events during the entire process cell nucleus is reprogrammed to an oocyte or egg pattern of gene expres- of reprogramming. sion. The second phase of reprogramming into new cell-types depends on We have recently revealed the importance of mesenchymal epithelial transi- the two fundamental mechanisms of development, namely the asymmetric tion as a critical event at the early phase of the reprogramming process as distribution of determinants to daughter cells and concentration-dependent well as the elasticity of the significant but early changes in cell properties. signalling. We have been investigating the genetic changes may be associated with the reprogramming process as well. In fact we were able to identify de novo generated copy number variations at the early phase of reprogramming, TET2 PROMOTES C/EBPA-INDUCED PRE-B which create a high level of genetic mosaicism in a single derived iPSC CELL TRANSDIFFERENTIATION BY line. Intriguingly, this genetic is decreasing when the cells are cultured for an intermediate period of time, due to selection against the highly mutant DE-REPRESSING MACROPHAGE GENES cellular compartment. We expect that our current genetic, epigenetic and Graf, Thomas, Kallin, Eric proteomic profiling of the reprogramming process will give us answers to pivotal questions essential for the future potential and use of iPSCs in hu- Center for Genomic Regulation and ICREA, Barcelona, Spain man medicine. Our previous work has shown that pre-B cells induced to transdifferenti- ate into macrophages by C/EBPa undergo extensive changes in gene expression, consisting in the down- or upregulation of thousands of genes (including B cell and macrophage specific genes). This happens within a matter of hours but it remained a mystery how genes become repressed or de-repressed. We have now found a novel mechanism for the de- repression of macrophage genes, which is mediated by Tet2. This gene encodes an enzyme that can hydroxylate methylated cytosines and plays a role in differentiation and cancer through a poorly understood mechanism. We discovered that Tet2 knockdowns repress the upregulation of specific

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Detailed Program and Abstracts – Friday, June 17

STEM CELLS, PLURIPOTENCY AND NUCLEAR Concurrent Session II Track A – Hematopoietic Stem Cells REPROGRAMMING Friday, June 17, 1:30 – 3:15 p.m. Jaenisch, Rudolf Whitehead Institute for Biomedical Research and M.I.T., Cambridge, MA, DEVELOPMENTAL MECHANISMS IN STEM USA CELLS AND CANCER The recent demonstration of in vitro reprogramming using transduction Reya, Tannishtha of 4 transcription factors by Yamanaka and colleagues represents a major Department of Pharmacology, University of California, San Diego, La Jolla, advance in the field. However, major questions regarding the mechanism CA, USA of in vitro reprogramming as well as the definition of pluripotent cell states need to be understood and will be one focus of the talk. Our work has focused on understand the signaling pathways that regulate the choice between stem cell renewal and commitment, and define how the a. Mechanisms of direct reprogramming: same signals are subverted in cancer. We have specifically studied develop- Direct reprogramming of somatic cells into induced pluripotent stem cells mental signals such as Wnt, Notch and Hedgehog which are critical regula- (iPSCs) can be achieved by over-expression of Oct4, Sox2, Klf4 and c-Myc tors of normal development in a variety of systems, and a major target of transcription factors, but only a minority of donor somatic cells can be re- mutation in human cancer. Our research using knockout and transgenic programmed to pluripotency. We have demonstrated that reprogramming is approaches suggests that these signals are activated in hematopoietic stem a continuous stochastic process where almost all donor cells eventually give cells, and that they functionally contribute to stem cell self-renewal in vivo. rise to iPSCs upon continued growth and transcription factor expression. In addition our data also shows that inhibition of these signals can block Inhibition of the p53/p21 pathway or over expression of Lin28 increased leukemia development and propagation in mouse models of the disease as the cell division rate and resulted in an accelerated kinetics of iPSC forma- well as in human leukemia cells. tion that was directly proportional to the increase in cell proliferation. These results suggest that the number of cell divisions is a key parameter driving epigenetic reprogramming to pluripotency. In contrast, Nanog over expression accelerated reprogramming in a predominantly cell division rate MULTI-POTENT HEMATOPOIETIC independent manner. PROGENITORS ARISE AT THE EXTRA- b. Different states of pluripotency: EMBRYONIC YOLK SAC PRIOR TO Human and mouse embryonic stem cells (ESCs) are derived from blastocyst stage embryos but have very different biological properties, and molecular HEMATOPOIETIC STEM CELL EMERGENCE IN analyses suggest that the pluripotent state of human ESCs isolated so far THE MOUSE EMBRYO corresponds to that of mouse derived epiblast stem cells (EpiSCs). We have Yoshimoto, Momoko, Porayette, Prashanth, Glosson, Nicole, rewired the identity of conventional human ESCs into a more immature state that extensively shares defining features with pluripotent mouse ESCs. Kaplan, Mark H., Yoder, Mervin C. This was achieved by exogenous induction of Oct4, Klf4 and Klf2 factors Pediatrics, Indiana University School of Medicine, Indianapolis, IN, USA combined with LIF and inhibitors of glycogen synthase kinase 3 (GSK3) and In the murine embryo, the first hematopoietic stem cells (HSC) that recon- mitogen-activated protein kinase (ERK) pathway. In contrast to convention- stitute lethally irradiated adult mice emerge at aorta-gonad-mesonephros al human ESCs, these epigenetically converted cells have growth properties, (AGM) region at E10.5. The emergence of lymphoid precursors capable of X chromosome activation state (aXa), a gene expression profile, and signal- forming mature T and B lymphocytes prior to this stage is still controversial. ing pathway dependence that are highly similar to that of mouse ESCs. The Cardiac contractions begin at E8.25, leading to the circulation of cells be- generation of “naïve” human ESCs will allow the molecular dissection of a tween various sites of hematopoiesis making it difficult to identify defini- previously undefined pluripotent state in humans, and may open up new tively the site where lymphoid precursors emerge. Hence a mouse model opportunities for patient-specific, disease-relevant research. was required to overcome the confounding factor of circulation so that the c. Gene targeting in human ES cells site of emergence of these lymphoid precursors can be identified. NCX1 A major impediment in realizing the potential of ES and iPS cells to study encodes for a sodium/calcium exchanger required for the initiation of a human diseases is the inefficiency of gene targeting. Using Zn finger heartbeat during embryogenesis. Ncx1 null embryos develop normally until mediated genome editing we have established efficient protocols to target E10.5, but die around E11.5 since circulation is never established in these expressed and silent genes in human ES and iPS cells. Finally, our progress in embryos. To determine the site of origin of hematopoietic precursors, E8.5 using iPS cells for therapy and for the study of complex human diseases will and 9.5 YS and para-aortic splanchnopleura (P-Sp) WT and Ncx1 null tis- be summarized. sues were digested into a single cell suspension and plated on OP9 stromal cells expressing delta-like protein 1 (OP9-DL1), which support T lympho- poiesis. Both Ncx1 null YS and P-Sp independently produced T cells includ- ing CD4 and CD8 double positive (DP), double negative (DN), and single positive (SP) cells with TCR β or γδ expression in vitro. Adoptive transfer of the lymphoid precursors derived from these sites into NOD/SCID/IL2Rg null neonatal mice revealed that they engrafted in all the lymphoid organs including thymus, spleen and liver in the recipient mice. These engrafted T cells were functional; they proliferated upon anti-CD3 stimulation and secreted IL2. Various kinds of mature T cells including naive T cells, memory T cell, regulatory T cells and NK T cells were detected in the recipient mice. Development of mature functional T lymphocytes from precursors isolated from YS and P-Sp much before the presence of a functional thymus brings forward the hypothesis for an extra-thymic origin of T cells in the early em- bryo which is different from adult lymphopoiesis. The presence of prospec- tive lymphoid precursors more than a day earlier than the adult reconsti- tuting HSCs first observed at E10.5 suggests that either the HSC may be

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Detailed Program and Abstracts – Friday, June 17

present at earlier points in development than is believed (and not detectable beneficial effect of glucose on hematopoiesis was conserved in adults: FACS by current assays) or that there may be a HSC independent emergence of analysis of kidney marrow following injury showed a more rapid recovery lymphoid precursors in the developing embryo. Future studies to examine of stem/progenitors after glucose exposure. Our data suggest that energy T cell development via lineage tracing studies will determine whether the T metabolism plays a key role in regulating HSC induction and expansion cell potential identified in the present work occurs in vivo. through ROS-mediated induction of hif1α and subsequent hematopoietic gene expression. These results could lead to novel therapeutic approaches for HSC modulation, and may unveil specific risks of obesity and diabetes METABOLISM-INDUCED REACTIVE OXYGEN for the fate and function of HSCs during gestation and in adults. SPECIES (ROS) AND HIF1α STIMULATION CONTROL THE INDUCTION AND EXPANSION TRACKING MURINE HEMATOPOIETIC STEM OF HEMATOPOIETIC STEM CELLS CELL DIFFERENTIATION IN VIVO WITH SINGLE Harris, James M.1, Harris, Lauren J.1, Cox, Andrew G.2, Cortes, CELL PRECISION: NEW INSIGHTS INTO THE Mauricio1, Garnaas, Maija K.2, Cutting, Claire C.2, Dovey, Michael CLONAL SUCCESSION VERSUS CLONAL 1 3 4 C. , Paw, Barry H. , Vander Heiden, Matthew G. , Goessling, STABILITY DEBATE Wolfram5, North, Trista E.1 1Department of Pathology, Beth Israel Deaconess Medical Center, Harvard Lu, Rong, Czechowicz, Agnieszka, Seita, Jun, Weissman, Irving L. Stem Cell Institute, Harvard Medical School, Boston, MA, USA, 2Genetics Stanford University, Stanford, CA, USA Division, Brigham and Women’s Hospital, Harvard Stem Cell Institute, Hematopoietic stem cells (HSCs) comprise a critical reservoir for maintaining Harvard Medical School, Boston, MA, USA, 3Hematology Division, blood and immune system homeostasis. Yet it remains unclear how HSCs Brigham and Women’s Hospital, Harvard Medical School, Boston, MA, are recruited for differentiation. The ‘clonal succession model’ proposes that USA, 4Koch Institute, Massachusetts Institute of Technology, Cambridge, small portions of the HSC reservoir sequentially commit to differentiation. MA, USA, 5Divisions of Genetics and Gastroenterology, Brigham and In contrast, the ‘clonal stability model’ suggests that a single static group Women’s Hospital, Harvard Stem Cell Institute, Harvard Medical School, of HSCs continuously replenish the blood supply throughout an organism’s Boston, MA, USA lifetime. Metabolic disorders, including obesity, diabetes, and their related complica- To discriminate between these two models, we have developed a novel tions have become the leading cause of preventable death in the U.S. While experimental system to track single HSC differentiation in vivo. Our effects of increased blood sugar on the cardiovascular system are estab- experimental system combines viral cellular labeling with high-throughput lished, consequences for the hematopoietic system are less characterized. sequencing to simultaneously track hundreds of cells with single cell resolu- Intriguingly, children born to mothers with gestational diabetes have a high- tion and sensitivity. We inserted unique 33-mer DNA codes into HSCs’ er risk of developing childhood leukemia, suggesting elevated glucose levels genomes using a lentivirus vector. ‘Barcoded’ HSCs were transplanted into may have lasting impact on hematopoiesis. To determine the consequence mice without any preconditioning regimens such as irradiation so as to of elevated glucose concentrations on hematopoietic stem cell (HSC) de- mimic natural physiological conditions. Five months post-transplantation, velopment, we treated zebrafish embryos with D-glucose during embryonic DNA codes from HSCs and several other hematopoietic populations were blood cell specification, from 12 to 36 hours post fertilization (hpf). HSCs extracted and analyzed using next-generation sequencing technology. in the Aorta-Gonad-Mesonephros (AGM) region were expanded in a dose-responsive manner, as assessed by in situ hybridization for runx1 and Our data suggests that all engrafted murine HSCs uniformly contribute to cmyb (n≥25-50 embryos/condition). L-glucose, the inactive enantiomer, did the blood supply after unconditioned transplantation. Our data do not sup- not affect HSCs, implying effects are independent of osmotic dynamics. A port the presence of quiescent HSCs nor the presence of exhausted HSCs. 3-fold enhancement in HSCs was observed after 1% glucose treatment by Clonal correlations between different stages of hematopoiesis indicate that qPCR for runx1, and FACS analysis of fluorescent HSC reporter embryos: in vivo differentiation is a linearly progressive development at the clonal runx1:eGFP, cmyb:eGFP and CD41:GFP; increased cellular proliferation level. We show that this naturally uniform differentiation can be per- (BrdU) was detected in the AGM, and elevated HSC number was sustained turbed by preparative regimens such as irradiation. We tracked the clonal without any apparent block in differentiation potential. HSC formation was contributions of HSCs to blood cells for over a year after various condition- impaired by morpholino knockdown of the glucose transporter glut1, and ing and non-conditioning regimens. We found that a static group of HSCs exposure to chemical inhibitors of glycolysis (lonidamine, ethyl-3-bromopy- continuously contribute to the blood supply between weeks 48 and 74 ruvate) and oxidative phosphorylation (cyanide, oxaloacetate) reversed post-transplantation. In contrast to the clonal succession model, our data the beneficial effects of glucose on HSCs. In contrast, pharmacological and suggests that all HSCs continuously replenish the blood supply over the genetic modulations of the metabolic endocrine hormones IGF and insulin lifetime of a mouse. did not alter the effects of glucose. These results suggest that glucose af- fects HSCs specifically through energy metabolism; the glycolytic intermedi- ate pyruvate similarly expanded HSCs. We postulated enhanced oxidative STRESS INDUCED ACTIVATION OF HSCS phosphorylation produced excess reactive oxygen species (ROS), which serve as hematopoietic signaling factors; treatment with the antioxidant Trumpp, Andreas1,2, Essers, Marieke A.G.1,2, Lutz, Raphael2, N-acetylcysteine (NAC) decreased HSC formation and could not be rescued Kern, Andrea3, Wurzer, Stephan3 by concomitant glucose exposure, while the oxidant 1-phenyl-2-thiourea 1Division of Stem Cells and Cancer, German Cancer Research Center expanded HSCs. Using the fluorescent ROS sensor dihydroethidium, we ob- (DKFZ), Heidelberg, Germany, 2HI-STEM gGmbH (Heidelberg Institute for served ROS in erythrocytes and CD41+ cells; exogenous glucose increased Stem Cell Technology and Experimental Medicine), Heidelberg, Germany, ROS levels. ROS are known inducers of the cellular hypoxia sensor, hypoxia 3German Cancer Research Center (DKFZ), Heidelberg, Germany inducible factor 1α (hif1α), which regulates expression of hematopoietic genes including vegf, epo, and inos, as well as the glucose transporter Maintenance of the blood system is dependent on dormant haematopoietic glut1. Chemical induction of hypoxia by cobalt chloride likewise enhanced stem cells (HSCs), which are characterized by multipotency and lifelong HSCs, and rescued the block in HSC induction following inhibition of energy self-renewal capacity. In order to both maintain a supply of mature blood metabolism or ROS. Likewise, by gene expression analysis, glucose expo- cells and not exhaust HSCs throughout the lifespan of the organism, most sure was observed to increase hif1α pathway associated transcripts. The HSCs remain deeply quiescent during adult homeostasis and only a limited 74 www.isscr.org Final Program

Detailed Program and Abstracts – Friday, June 17 number are cycling at any given time. Omics technologies have been ap- MAMMALIAN GENES INDUCE IPS-LIKE CELLS plied to characterize the different HSC populations and the various molecu- lar programs that are active will be discussed. IN NON-MAMMALIAN SPECIES 1 1 1 The balance between self-renewal and differentiation of HSCs is controlled Rossello, Ricardo A. , Chen, Chun-Chun , Howard, Jason T. , by external factors such as chemokines and cytokines, as well as interac- Dai, Rui2, Hochgeschwender, Ute2, Jarvis, Erich D.1 tions of HSCs with its niche environment. We have recently shown that the 1Neurobiology, Duke University, Howard Hughes Medical Institute, cytokine IFNα very efficiently activates dormant HSCs in vivo. Within hours Durham, NC, USA, 2Duke University, Durham, NC, USA after treatment of mice with IFNα, HSCs exit G and enter an active cell cycle. 0 Cells are fundamental units of life, but little is known about evolution of In general, IFNα is produced in response to viral infections by cells of the cell states. Induced pluripotent stem cells (iPSCs) are once differentiated immune system, and plays an important role in the antiviral host defense. We cells that have been reprogrammed to an embryonic stem cell (ESC) state, then asked whether endogenous IFNα is also produced in response to other providing a powerful platform for biology and medicine. So far, induction types of bone marrow stress and whether this affects the proliferation rate has been limited to a few mammalian species. However, important issues in of HSCs. To monitor IFNα production in vivo, we have generated a reporter biology are addressed in experimental systems other than mammals, such mouse strain and found that treatment with both the chemotherapeutic as birds, fish and flies. Some of these animals have traits similar to humans, agent 5-FU as well as the endotoxin LPS leads to the production of IFNα such as vocal learning in songbirds, that are not found in closely related in the vicinity of HSC/progenitors. In addition, LPS treatment in vivo also non-human primates or commonly used laboratory animals. For some of induced a strong increase in HSC cycling. Surprisingly, this does not require these systems, stem cells have not been isolated or are hard to generate. IFNAR signaling as mice lacking the IFNα receptor still respond to LPS. LPS Here we report that the same four transcription factor genes can reprogram binds and signals via the TLR4-CD14 receptor complex and HSCs of mice differentiated cells to an iPSC state in other vertebrate and invertebrate lacking TLR4 are no longer activated by LPS. Strikingly, LPS induced HSC species spanning over 550 million years removed from a common ancestor. activation is not direct, but mediated by a myeloid cell type and correlates This includes birds (galliforms and songbirds), fish (zebrafish), and an insect with increased cell surface expression of Sca-1 on HSCs, similar to what oc- (drosophila). Moreover, the mammalian (mouse and human) homolog of curs upon IFNα treatment. Moreover, as for IFNα, the up-regulation of Sca-1 these genes reprogrammed the non-mammalian cells of all species tested. is required for LPS induced proliferation, since Sca-1-/- mice fail to respond to The transfected vertebrate cells exhibited features of mammalian iPSCs. LPS stimulation. In summary, these data suggest that not only virus induced Control fibroblasts transduced with a GFP only construct and seeded on IFNα, but also LPS of gram negative bacteria induces cycling of stem/progen- stem cell media did not show any of these features for all species. Five days itors and dormant HSCs in a Sca-1 dependent manner. Finally, since activated after the first passage, the iPSC-like cells formed characteristic clusters, HSCs become sensitized to chemotherapy, a sequential strategy in which cells which were not evident in the control fibroblast group. The transduced cell are activated prior to standard or targeted chemotherapy may be developed clusters stained for alkaline phosphatase activity. After 20 days, species- to eliminate otherwise resistant dormant leukemic stem cells. specific endogenous homologs of the 4 mouse genes and endogenous Nanog and Vasa stem cell marker genes were overexpressed in most species, indicating reprogramming. Proliferation was also enhanced and Concurrent Session II Track B – telomerase activity was present, indicating a self-renewal reprogrammed state had been achieved. We can propagate the cells, so far, up to the 7th Stem Cells in Non-Mammalian Models passage with these characteristics. Pluripotency was achieved by generat- ing embryoid bodies (EBs) for each of the transduced cells lines, which did Friday, June 17, 1:30 – 3:15 p.m. not occur with control fibroblasts. These had a characteristic morphology, typical of EBs, and expressed gene markers for all three germ layers. These DEFINITION AND DYNAMICS OF THE CARDIAC markers were not observed in the control fibroblast cells or in the undif- PROGENITOR POOL IN ZEBRAFISH ferentiated iPSC-like cells. Lastly, we were able to demonstrate in-vivo incorporation of these cells, by producing an embryonic chimeric chicken Yelon, Debbie with the labeled iPSC-like cells. These findings are the first we are aware of Division of Biological Sciences, University of California, San Diego, La to generate iPS-like cells for commonly used non-mammalian animal mod- Jolla, CA, USA els, to show cross-lineage stem cell induction, and to generate stem cells for songbirds and drosophila, despite years of effort. The findings suggest that Protocols for creating multipotent cardiovascular progenitor cells (CPCs) in the stem-cell state is highly conserved across non-mammalian vertebrates vitro are continually improving in their efficiency. However, it is less clear and invertebrates, lending new insights into the nature of stem cells and cell how to control the differentiation of CPCs into cardiomyocytes in a robust evolution. and scalable fashion. To identify molecules that trigger the decision to dif- ferentiate, we study CPC development in zebrafish, where we can track the progression of myocardial differentiation in vivo. The embryonic zebrafish heart is built through two distinct phases of cardiomyocyte differentiation: THE EPIGENEETIC REGULATION OF RENAL an initial phase that establishes the heart tube and a later phase that recruits STEM CELL NUMBER IN ZEBRAFISH cardiomyocytes to create the outflow tract (OFT). Fgf signaling is required 1 2 3 3 for OFT formation, but it is unknown which genes function downstream Ma, Dongdong , Diep, Cuong , de Groh, Eric , Hukriede, Neil , 2 4 of Fgf signaling to execute the production of cardiomyocytes. Through Alan, Davidson , Handin, Robert I. gene expression profiling, we have identified a cell adhesion molecule, 1Hematology Division, Brigham & Women’s Hospital, Boston, MA, USA, cell adhesion molecule 4 (cadm4), that is repressed by Fgf signaling and 2Center for Regenerative Medicine, Massachusetts General Hospital, plays a crucial role in inhibiting CPC differentiation. Strikingly, knockdown Boston, MA, USA, 3Microbiology and Molecular Genetics, University of of cadm4 function results in a dramatic enhancement of the number of Pittsburgh, Pittsburgh, PA, USA, 4Brigham & Women’s Hospital, Boston, OFT cardiomyocytes. Our results suggest a model in which Fgf signaling MA, USA promotes cardiomyocyte recruitment by limiting cadm4 expression and We have recently reported the identification and characterization of a pop- thereby altering the cell adhesion properties of CPCs. Thus, we propose a ulation of renal progenitor cells, which can form fully functional nephrons in novel adhesion-based mechanism for the regulation of cardiomyocyte dif- zebrafish kidney. The studies were carried out by injecting clusters of 10-30 ferentiation. lhx1+ positive cells derived from the whole kidney marrow of a double transgenic zebrafish strain--Tg(lhx1:GFP,cdh17:mCherry) into the head 75 ISSCR 9th Annual Meeting www.isscr.org

Detailed Program and Abstracts – Friday, June 17

kidney of irradiated recipient fish. The development of mCherry positive S1P-dependent HSPC trafficking. We found that this receptor is expressed nephrons after the injection of GFP+cells provided strong evidence of neph- in the dorsal aorta and CHT, and that morpholino knockdown of s1pr1 ron formation from transplanted progenitors. We now report that lhxl+ cell resulted in reduction of hematopoietic markers in the CHT. We could also clusterss can be dissociated, the single cells isolated by flow cytometry and reproduce this result using a highly specific s1pr1 receptor antagonist injected into the cardiac circulation of irradiated recipients where they home (W146), or by depletion of endogenous S1P using a sphingosine kinase to the kidney, migrate into the renal interstitium and form cdh17:mCherry+ inhibitor (SKI-2). Previous studies showed that UV uncaging of fluores- nephrons. Engraftment and new nephron formation can be achieved with cent lineage tracer specifically in the dorsal aorta marks the HSPCs that as few as 10 lhx1+ cells. The number of mCherry+ nephrons increases after subsequently seed the CHT. We repeated this experiment in the presence treatment of transplanted fish with nephrotoxic doses of gentamycin. lhx1+ of s1pr1 receptor antagonist and found that HSPCs still emerge from the cells transplanted into secondary hosts also form nephrons and persist in dorsal aorta and undergo homing to the CHT, but then appear to lose irradiated host fish for as long as 18 months after transplantation. We next their progenitor-like morphology and differentiate. We examined this more sought to study the regulation of stem cell number within the kidney. We closely by using transgenic over-expression of dominant negative s1pr1 exposed Tg(lxh1:GFP) fish to nephrotoxic doses of gentamycin and then specifically in HSPCs, followed by time-lapse live imaging, and found that isolated kidneys from treated and control fish. We noted by flow cytometry these HSPCs had difficulty engrafting in the CHT. Given previous reports that 0.01% of cells from transgenic donors were GFP+ in control fish. The that cxcr4 and S1P pathways could interact during homing of adult HSPCs, number of GFP+ cells went up within 2-3 days of gentamycin treatment, we decided to test for possible synergism between these signals in the peaked at 7 days and then gradually returned to baseline levels 28 days embryo. We used combination chemical genetics to apply dose matrices of after exposure to gentamycin. Since we have previously reported that a two different chemicals to modulate the cxcr4 and S1P pathways simultane- histone deacetylase inhibitor, 4-methyl-phenyl-thio-butadione (4-mPTB), ously. We found specific dosage ratios that could synergistically increase or increased the pronephric field in zebrafish embryos, we injected fish with decrease hematopoiesis in the CHT, demonstrating an interaction between varying doses of 4-mPTB. In contrast to gentamycin, 4-mPTB was not cxcr4 and S1P signaling. Together, this chemical and genetic data highlights nephrotoxic. However, a single injection increased the number of GFP+ cells a novel role for S1P during migration of HSPCs between successive niches three fold and three injections on alternate days increased the number of in the embryo. GFP+ cells on day 7 eight fold. Finally, we observed that 4-mPTB aug- mented the already robust response to gentamycin and increased the final number of GFP+ cells produced by renal injury. We believe that the lhx1+ INTESTINAL RENEWAL AND REGENERATION IN cells we have isolated from zebrafish kidney fulfill the usually accepted criteria for adult stem cells and account for the remarkable regenerative THE PLANARIAN SCHMIDTEA MEDITERRANEA ability of zebrafish kidneys. We now plan to test the effect of 4-mPTB and Newmark, Phillip A., Forsthoefel, David, James, Noelle, Waters, other HDAC inhibitors on renal stem cell number and nephron production Forrest, Park, Amanda, Escobar, David, Stary, Joel in zebrafish under various experimental conditions and, eventually, extend the studies to fetal and neonatal mice. HHMI/University of Illinois at Urbana-Champaign, Urbana, IL, USA In planarians, pluripotent somatic stem cells called neoblasts replenish cells lost to normal physiological turnover and regenerate tissues in response TRAFFICKING OF ZEBRAFISH HEMATOPOIETIC to injury. Utilizing the model planarian Schmidtea mediterranea, we have characterized renewal and repair of the intestine, the organ responsible for STEM CELLS DURING EMBRYONIC digestion and nutrient distribution. In contrast to organisms that possess DEVELOPMENT resident intestinal stem cells, we demonstrate that in planarians, intestinal epithelial cells arise as the progeny of neoblasts, differentiating along the Tamplin, Owen J., Zon, Leonard I. entire length of the intestine in intact animals, but mainly near the site of Hematology/Oncology, Children’s Hospital Boston, Boston, MA, USA amputation during regeneration. We also report evidence that differenti- Hematopoietic stem and progenitor cells (HSPCs) self-renew, give rise to ated intestinal tissue remodels as polarity and symmetry are restored in all blood cell types, and replenish blood throughout adulthood. Definitive amputated tissue fragments. We have developed a novel method for the HSPCs first arise from the hemogenic endothelium of the dorsal aorta, are purification of intestinal phagocytes (the absorptive epithelial cell type), released into circulation, and then seed the fetal liver as an intermediate enabling microarray-based identification of over 1000 genes with gut-en- tissue before colonizing the adult bone marrow. Although adult trafficking riched expression. An RNA interference screen has identified genes required of HSPCs has been extensively studied, little is known about the signals that for intestinal epithelial cell differentiation, maintenance of normal intestinal regulate HSPC migration during development. In zebrafish, the first HSPCs branch morphology, and neoblast proliferation. The latter result suggests also emerge from the dorsal aorta, but the intermediate tissue they migrate that the intestine may play a niche-like role in the regulation of neoblast to is the caudal hematopoietic tissue (CHT), a vascular plexus in the ventral dynamics. Investigation of intestinal regeneration in planarians will help tail of the embryo. We wanted to find small molecules that could affect the reveal evolutionarily conserved mechanisms of stem cell-based renewal and signaling pathways involved in migration of HSPCs to the CHT. First, we repair of digestive organs. chose the cxcr4-cxcl12 signaling axis as a candidate pathway based on its known role in adult HSPC homing and engraftment, and our expression studies of cxcr4 and cxcl12 orthologs in the zebrafish CHT. A cxcr4 antago- nist AMD3100 dose-dependently reduced expression of hematopoietic progenitor markers cmyb and runx1 in the CHT, suggesting this receptor is required for HSPC seeding of the CHT. Next, we performed a chemical genetic screen using a library of 2400 known bioactive small molecules for effects on CHT homing. Three primary classes of small molecules were discovered: 1) those that increase HSPC marker expression in the CHT; 2) those that decrease it; 3) and those that change the pattern of expression. Surprisingly, sphingosine-1-phosphate (S1P), a signal known to regulate adult progenitor and lymphocyte trafficking, altered the expression pattern of HSPC markers in the embryo. We examined the embryonic expression and function of s1pr1, the G protein-coupled receptor that is important for

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Detailed Program and Abstracts – Friday, June 17

required for maintenance of quiescent HSCs in bone marrow niche and that Concurrent Session II Track C – Stem Cell Asymmetry it anchors Frizzled8 protein between HSCs and niche cells, thus ensuring Friday, June 17, 1:30 – 3:15 p.m HSCs receive niche emanated non-canonical Wnt signaling. ASYMMETRIC CELL DIVISION AND TUMORIGENESIS IN NEURAL STEM CELL ASYMMETRIC CELL DIVISIONS PROMOTE LINEAGES NOTCH-DEPENDENT EPIDERMAL DIFFERENTIATION Knoblich, Juergen A.1, Neumüller, Ralph A.1, Richter, Constance1, Fischer, Anja1, Novatchkova, Maria2, Neumüller, Klaus Williams, Scott E., Beronja, Slobodan, Fuchs, Elaine G.1 Mammalian Cell Biology & Development, The Rockefeller University, New 1IMBA – Institute of Molecular Biotechnology, Vienna, Austria, 2IMP, York, NY, USA Vienna, Austria Stem and progenitor cells utilize asymmetric cell divisions to balance pro- Stem cells are characterized by their ability to generate both self-renewing liferation and differentiation. Evidence from invertebrates shows that this and differentiating daughter cells. We are using Drosophila and mouse process is regulated by proteins asymmetrically distributed at the cell cortex neural stem cells as models to understand how these differences are during mitosis: Par3-Par6-aPKC, which confer polarity, and Gαi-LGN/ established and regulated. Drosophila neural stem cells called neuroblasts AGS3-NuMA-dynein/dynactin, which govern spindle positioning. Here we undergo repeated rounds of asymmetric cell division during which they focus on developing mouse skin, where progenitor cells execute a switch segregate the cell fate determinants Numb, Prospero and Brat into one of from symmetric to predominantly asymmetric divisions concomitant with the two daughter cells. In this cell, the determinants prevent self-renewal stratification. Using in vivo skin-specific lentiviral RNAi, we investigate and induce differentiation. When one or more determinants are missing, spindle orientation regulation and provide direct evidence that LGN (also all daughter cells continue to proliferate leading to the formation of a stem called Gpsm2), NuMA and dynactin (Dctn1) are involved. In compromising cell derived brain tumor. How determinants segregate asymmetrically, how asymmetric cell divisions, we uncover profound defects in epidermal strati- they prevent self-renewal and how stem cell tumors are formed are the key fication, differentiation and barrier formation, and implicate Notch signaling questions we are trying to answer.For this, we have used a library of 20,000 as an important effector. Our study demonstrates the efficacy of applying transgenic RNAi lines to screen for genes involved in self-renewal control. RNAi in vivo to mammalian systems, and the ease of uncovering complex We have identified over 600 genes required in neuroblasts and have quanti- genetic interactions, here to gain insights into how changes in spindle fied their RNAi phenotypes to assign particular biological functions. Using orientation are coupled to establishing proper tissue architecture during skin hierarchical clustering of quantitative phenotypic data and integration with development. protein interaction data, we have created regulatory networks for neural stem cell self-renewal, differentiation and tumorigenesis and identified a set of transcriptional regulators essential for self-renewal. Analysis of the genes CHROMOSOME STRAND SEGREGATION in those networks has revealed a surprising role for duplicated ribosomal DURING DROSOPHILA MALE GERMLINE STEM proteins in self-renewal control. In addition, our data reveal that alternative splicing of transcriptional regulators, chromatin remodeling and the regula- CELL DIVISION tion of transcriptional elongation are important for controlling self-renewal Yadlapalli, Swathi, Yamashita, Yukiko and differentiation and preventing tumor formation in stem cell lineages. University of Michigan, Ann Arbor, MI, USA Throughout the life of an organism, stem cells are required to proliferate A ROLE OF NON-CANONICAL WNT SIGNALING and supply differentiated cells while avoiding the potentially deleterious effects of DNA mutations resulting from repeated cell cycles. It has been IN MAINTAINING HEMATOPOIETIC STEM CELLS hypothesized that stem cells might be accomplishing this remarkable feat by Sugimura, Ryohichi, He, Xi, Venkatraman, Aparna, Perry, John, retaining older (“immortal”) DNA strands during asymmetric cell divisions, Li, Linheng thereby excluding all replication-induced mutations into the differentiating daughters (Immortal Strand Hypothesis - ISH). In addition, other models Stowers Institute, Kansas City, MO, USA have also been proposed in which stem cells asymmetrically segregate only The signaling and the related molecular mechanism underlying hematopoi- a subset of chromosomes for different reasons such as retention of epigen- etic stem cell (HSC) maintenance is a fundamental question. Cadherin EGF etic memories. Recently, the idea has emerged, provoked by a finding from LAG seven-pass G-type receptor2 (Celsr2) is a member of the Flamingo our laboratory that Drosophila male germline stem cells consistently inherit gene family that mediates non-canonical Wnt signaling. From our previ- the mother centrosome, that the mother centrosome might be utilized ous microarray screening, we observed that Celsr2 was predominantly as a means of asymmetrically segregating DNA strands. In order to test if expressed in HSCs compared with hematopoietic progenitors. Since non- germline stem cells (GSCs) follow the immortal strand model, we scored canonical Wnt signaling has been implicated to maintain quiescent adult the outcome of DNA strand segregation by using 5-bromo-2-deoxyuridine tissue stem cells, we hypothesize that Celsr2 may maintain HSCs through labeling combined with direct visualization of GSC-gonialblast (differentiat- non-canonical Wnt signaling. In our transplantation model, knocking down ing daughter) pairs. Our data unambiguously demonstrate that Drosophila via lentivirus-mediated shRNA expression of Celsr2 decreased long-term male GSCs do not follow the immortal strand model. We are currently hematopoietic reconstitution. HSCs in bone marrow were reduced and probing the possibility whether GSCs might be segregating only a subset lost quiescence. Immunostaining showed that Celsr2 protein was localized of chromosomes asymmetrically. To this end, we recently adapted the CO- between HSCs and different niche components, including osteblastic-lining FISH (chromosome orientation fluorescence in situ hybridization) technique cells, megakaryocytes, and endothelial cells. Furthermore, we also found to follow the segregation of template DNA strands in vivo. Briefly, the that a non- canonical Wnt receptor, Frizzled8 protein, was expressed in CO-FISH method involves 1) removal of newly formed strand by creating HSCs and co-localized with Celsr2 protein. Celsr2 was required for focal nicks at the sites of BrdU incorporation and treating with exonuclease, 2) distribution of Frizzled8 protein on the side of HSCs contacting with the hybridization of single stranded template DNA to chromosome-specific OP9 cells in vitro osteoblastic-lining cells in vivo. We propose that Celsr2 is satellite repetitive probes. Strikingly, our preliminary study indicates that

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Detailed Program and Abstracts – Friday, June 17

GSCs preferentially inherit a certain chromosomal strand of Y chromosome. MITOTIC CHRONOLOGIES IN MURINE GUT We are currently probing potential asymmetries in the segregation of other chromosomes. PROGENITORS Kozar, Sarah, Houghton, Carol, Kemp, Richard, Winton, Doug Cambridge Research Institute, Cambridge, United Kingdom NEURAL STEM AND PROGENITOR CELLS — The cells of the intestinal epithelium are characterized by a high rate of A CELL BIOLOGICAL AND EVOLUTIONARY turnover and are maintained by a population of stem cells located at the PERSPECTIVE base of each intestinal crypt. A number of markers have been applied to identify stem cells in the intestine, including Bmi1 and Lgr5. However Huttner, Wieland B. cells expressing these markers occupy different cellular positions and have Max Planck Institute of Molecular Cell Biology and Genetics, Dresden, different cell cycle characteristics. It has recently been demonstrated that Germany the intestinal epithelium is maintained by a population of equipotent stem cells. Thus all stem cells, regardless of marker expression, must contribute Our group studies the cell biological mechanisms of neurogenesis in the equally to the same stem cell pool. Therefore, what are the implications developing cerebral cortex in the context of mammalian brain evolution, of heterogeneity in cell cycle times within this population? To answer this specifically the proliferation versus differentiation of neural stem and pro- question we have developed a mitotic clock that uses a novel clonal report- genitors cells. In the developing rodent cortex, two principal classes of these ing strategy to determine the cumulative number of stem cell divisions cells can be distinguished, (i) progenitors dividing at the ventricular, i.e. undergone in the intestine over time. In this system, reporter expression is apical, surface of the ventricular zone (VZ), called apical progenitors (APs; caused by mutations acquired in a dinucleotide repeat. Because intestinal neuroepithelial cells and radial glial cells), and (ii) progenitors dividing in a crypts are monoclonal units, they effectively record their cellular history. more basal, abventricular location, notably the subventricular zone (SVZ), Hence, characteristics of stem cell behavior are implied from the pattern of called basal progenitors (BPs, also called intermediate progenitors). Focus- reporter expression within the crypts. With this model we have observed ing on the embryonic mouse cortex, we have been studying the following significant differences in the rate of stem cell division in wild type small issues: intestine and colon. We are also investigating changes in stem cell behavior (1) the neurogenic lineage from APs and BPs to neurons, and the underly- in the hyperproliferative intestines of p27 deficient mice. Finally this system ing molecular machinery, using the marker Tis21; (2) the role and ma- should provide insights into potential changes in the background mutagen- chinery of interkinetic nuclear migration and delamination, including the esis and tumour age in Apc Min mice. primary cilium and centrosome; (3) apical-basal cell polarity, cleavage plane orientation and symmetric versus asymmetric division of APs and BPs; (4) the role of the microcephaly gene Aspm in symmetric AP divisions and its IDENTIFICATION OF MAMMARY CANCER more general role in organ size; (5) apical membrane constituents, notably the cholesterol binding protein prominin-1/CD133; (6) prominin-1-bearing STEM CELLS USING TG11.5KB-GFP MICE extracellular membrane particles relased into the ventricular fluid from the Bai, Lixia, Rohrschneider, Larry R. midbody and primary cilium of APs, and their role in differentiation; (7) the basal process of APs in mitosis; (8) the role of cell cycle length in stem Basic Sciences, Fred Hutchinson Cancer Research, Seattle, WA, USA and progenitor cell proliferation versus differentiation. We have recently Although the existence of cancer stem cells in mammary tumors has extended our investigations from the lissencephalic mouse model to spe- been proposed, the exact identity of such cells remains unknown due to cies developing a gyrencephalic cortex. We find that progenitors in the lack of stem cell specific markers. A novel shorter SH2-containing inositol outer SVZ (OSVZ) of human and ferret retain certain features of radial glia, 5’-phosphatase (SHIP) isoform called s-SHIP has been identified specifi- notably a basal process contacting the basal lamina. This epithelial feature cally in embryonic stem cells and adult hematopoietic stem cells, but not appears to allow integrin-mediated, repeated asymmetric divisions of OSVZ in differentiated cells. We previously demonstrated that green fluorescent progenitors, which provides a basis for neocortical expansion. protein positive (GFP+) cap cells at puberty and basal alveolar bud cells at pregnancy are stem cells of the mammary gland using the transgenic mice termed Tg11.5kb-GFP, in which enhanced GFP expression is driven by the 11.5kb s-SHIP promoter. Based on these data, we postulate that GFP Concurrent Session II Track D – Epithelial Stem Cells (s-SHIP) is likely to be expressed specifically in cancer stem cells during mammary tumorigenesis. To test our hypothesis, we crossed our Tg11.5kb- Friday, June 17, 1:30 – 3:15 p.m. GFP mice with two breast tumor mouse models: TgMMTV-Wnt1 and RECIPROCAL SIGNALLING BETWEEN TgMMTV-ErbB2. We examined GFP expression in mammary tumors from Tg11.5kb-GFP;MMTV-Wnt1 mice and Tg11.5kb-GFP;MMTV-ErbB2 mice, EPIDERMAL STEM CELLS AND CELLS IN THE characterized GFP+ cells by immunofluorescence and flow cytometry, and NICHE further analyzed the stem cell activity of GFP+ mammary tumor cells by lim- ited dilution and serial transplantation. We also performed DNA microarray Watt, Fiona M. analysis. We found that GFP expression is turned on in basal cells of Wnt1 CRUK Cambridge Research Institute, Cambridge University, Cambridge, oncogene caused tumors, which contain both basal/myoepithelial cells and United Kingdom luminal epithelial cells. GFP was not expressed in ErbB2 induced tumors, which are composed of only luminal cells. We also analyzed GFP+ cap cells Stem cell behaviour is controlled both by intrinsic mechanisms and by exter- in 5 week-old mammary glands of Tg11.5kb-GFP;MMTV-Wnt1 mice and nal signals from the local microenvironment or niche. Interactions with the Tg11.5kb-GFP;MMTV-ErbB2 mice, respectively. Tumors have not formed niche are reciprocal, since stem cells are capable of remodeling their environ- yet in 5 week-old Wnt1+ or ErbB2+ mammary glands; however, compared ment. Using adult epidermis as an experimental model, my lab has been in- to 11.5kb-GFP+/-MMTV-Wnt1-/- litermate mammary glands, the percent- vestigating the relative importance of specific intrinsic and extrinsic signals in age of GFP+ cap cells in puberty 11.5kb-GFP+/-MMTV-Wnt1+/- glands regulating stem cell fate. We have also discovered how altered intrinsic signal- increased about two-fold, 5.64% vs 17.25%. The percentage of GFP+ cap ing in stem cells impacts on neighbouring cells of the dermis. These findings cells in puberty 11.5kb-GFP+/-MMTV-ErbB2+/- glands was similar to that of are likely to be relevant to understanding how oncogenic changes in cells of 11.5kb-GFP+/-MMTV-ErbB2-/- littermate glands, 5.23% vs 5.14%. GFP+ multi-layered epithelia cause changes in the underlying tumour stroma. epithelial cells in Wnt1 tumors are stained positive with basal cell markers 78 www.isscr.org Final Program

Detailed Program and Abstracts – Friday, June 17 such as smooth muscle actin (SMA) and 14 (weak), and negative ADIPOCYTES REGULATE THE SKIN STEM CELL with luminal cell marker E-cadherin. GFP+ Wnt1 tumor cells are CD24+, CD29hi, CD49fhi, Sca-1low, CD133+. And most GFP+ tumor cells are Ki67+. NICHE Upon transplantation, basal GFP+ tumor cells can continuously regenerate Horsley, Valerie1, Festa, Eric1, Fretz, Jackie2, Berry, Ryan3, new tumors with high efficiency, and give rise to both basal/myoepithe- Schmidt, Barbara3, Rodeheffer, Matthew4, Horowitz, Mark2 lial and luminal epithelial cells in tumor outgrowths. In addition, our high 1Departments of Molecular, Cell and Developmental Biology, Yale Stem quality gene expression data may identify genes and signaling pathways Cell Center, New Haven, CT, USA, 2Department of Orthopædics and that regulate stem cells and breast tumor formation. In summary, GFP+ Rehabilitation, New Haven, CT, USA, 3Molecular Cell Biology, Genetics tumor cells from Tg11.5kb-GFP;MMTV-Wnt1 mice not only self-renew but and Development Program, New Haven, CT, USA, 4Section of Comparative also differentiate, and represent activated cancer stem cells during mam- Medicine, Yale Stem Cell Center, New Haven, CT, USA mary tumor formation and growth. The expansion of GFP+ cap cells (stem cells) in puberty 11.5kb-GFP+/-MMTV-Wnt1+/- mammary glands suggests Stem cells in the skin maintain tissue homeostasis and regeneration through that mammary stem cells are likely the targets of transformation by Wnt1 dynamic interactions with multiple cell types in the underlying dermis, oncogene. including fibroblasts, blood vessels, and adipocytes. The functional role of many of these cellular interactions in the skin is not well understood. We have analyzed the biology and function of subdermal adipocytes during YAP1 ACTS DOWNSTREAM OF ALPHA- skin tissue homeostasis. Subdermal adipocytes compose a unique white adipose tissue depot that underlies the dermal fibroblasts and surrounds the CATENIN TO CONTROL EPIDERMAL hair follicle during hair growth. Our data show that a critical interplay exists PROLIFERATION between hair follicle stem cells and subdermal adipocytes. During murine

1 1 2 follicular regeneration and stem cell activation, adipogenesis characterizes Mohseni, Morvarid , Schlegelmilch, Karin , Kirak, Oktay , changes in the skin tissue microenvironment through de novo formation 3 4 5 Pruszak, Jan , Zhou, Dawang , Vasioukhin, Valera , Avruch, of subdermal adipocytes. We find that subdermal adipocytes modulate Joseph4, Brummelkamp, Thijn6, Camargo, Fernando1 the activity of murine adult follicular stem cells. Furthermore, we implicate 1Stem Cell and Regenerative Biology/Stem Cell Program, Harvard signaling molecules that regulate these processes. These data suggest that University/Children’s Hospital, Boston, Cambridge/Boston, MA, USA, adipocytes are positive regulators of the stem cell niche in the skin and 2Whitehead Institute for Biomedical Research, Boston, MA, USA, 3Harvard define a vital crosstalk between adipocytes and adult epithelial stem cells in Stem Cell Institute/Whitehead Institute for Biomedical Research,, murine skin. Cambridge, MA, USA, 4Department of Molecular and Cellular Biology, Massachusetts General Hospital, Boston, MA, USA, 5Fred Hutchinson Cancer Research Center, Seattle, WA, USA, 6Whitehead Institute for Biomedical Research, Cambridge, MA, USA Concurrent Session II Track E – Endodermal Stem Cells During mammalian development, the proliferation of tissue-specific pro- Friday, June 1, 1:30 – 3:15 p.m. genitor and stem cells needs to be tightly regulated to produce organs of a predetermined size. These regulatory mechanisms also modulate homeo- STEM CELL APPROACHES TO DISSECT LUNG stasis of adult tissues and regenerative processes. While several signaling CANCER BIOLOGY molecules have been implicated in controlling stem cell proliferation, little is known about the endogenous mechanisms that ‘sense’ or provide informa- Kim, Carla F. tion about organ size. The Hippo signaling pathway is a potent mechanism Children’s Hospital Boston & Harvard Medical School, Boston, MA, USA that restricts tissue size by limiting cell proliferation and promoting apop- tosis. Here we use gain- and loss-of-function models in mice to show that Lung injury models have suggested that each niche in the lung contains Yap1 is an essential regulator of epidermal maintenance and stem cell prolif- its own stem or progenitor cell population. We previously determined that erative capacity. The restricted activation of Yap1 in K14-positive epider- cells expressing the alveolar type II (AT2) cell marker, SPC, and the Clara cell mal progenitors in mice leads to the expansion of the epidermal stem cell marker, CCSP, are present in normal lung, and that they constitute a lung compartment and squamous-cell carcinoma-like tumors. Moreover, the loss stem cell population named bronchioalveolar stem cells (BASCs). BASCs can of Yap1 results in impaired epidermal proliferation and the failure of skin be isolated using fluorescence activated cell sorting (FACS) methodology expansion. These data suggest that levels of active Yap1 are a critical and based on the presence of the surface marker Sca1, self-renew over multiple physiological determinant of epidermal stem cell proliferative capacity. In passages, give rise to bronchiolar and alveolar cells in culture, and prolifer- addition, we identify alpha-catenin, a known tumor suppressor in the skin, ate in response to lung injury. We hypothesize that BASCs execute repair as a crucial upstream negative regulator of Yap1 in epidermal cells in vitro after bronchiolar and alveolar cell injury in vivo, while other bronchiolar and and in vivo. This work makes a direct connection among the environmental alveolar progenitor cells maintain lung homeostasis. Current data from our signaling cues, such as cell-density, and a transcriptional regulator, and thus lab and others suggest that Clara cells, AT2 cells, or other unknown cells, in provides a mechanistic paradigm for the regulation of stem cell proliferation addition to BASCs, are lung stem cells. This talk will focus on how our lab and tissue expansion in response to extracellular cues. is seeking to elucidate the biology of these important lung cells. First, I will describe our work to introduce lung stem cells in vivo for functional tests. Second, I will discuss new genetically engineered mice we have created to determine the potential of BASCs and other lung epithelial cells. Finally, I will share our efforts to identify additional markers to purify BASCs and determine the mechanisms that regulate lung stem cells.

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Detailed Program and Abstracts – Friday, June 17

A NOVEL STEM/PROGENITOR CELL TGFβ FAMILY MEMBERS AND WNT SIGNALING POPULATION FROM MURINE TRACHEAL REGULATE PANCREATIC SPECIFICATION OF SUBMUCOSAL GLAND DUCTS WITH HUMAN PLURIPOTENT STEM CELLS MULTIPOTENT REGENERATIVE POTENTIAL Nostro, M. Cristina1, Sarangi, Farida1, Ogawa, Shinichiro1, Hegab, Ahmed E.1, Ha, Vi Luan1, Gilbert, Jennifer L.1, Zhang, Holtzinger, Audrey1, Corneo, Barbara2, Li, Xueling3, Micallef, Kelvin X.2, Malkoski, Stephen P.3, Chon, Andy T.1, Darmawan, Suzanne J.3, Park, In-Hyun4, Daley, George Q.5, Elefanty, Andrew Daphne O.1, Bisht, Bharti1, Ooi, Aik T.1, Pellegrini, Matteo4, G.3, Stanley, Edouard G.3, Keller, Gordon1 Nickerson, Derek W.1, Gomperts, Brigitte N.1 1McEwen Centre for Regenerative Medicine, Toronto, ON, Canada, 2 3 1Pediatrics, Mattel Children’s Hospital UCLA, Los Angeles, CA, USA, New York Neural Stem Cell Laboratories, Rensselaer, NY, USA, Monash 4 2Biological Chemistry, UCLA, Los Angeles, CA, USA, 3Division of Immunology and Stem Cell Laboratories, Clayton, Australia, Yale Stem 5 Pulmonary Sciences and Critical Care Medicine, University of Colorado, Cell Center, New Haven, CT, USA, Children’s Hospital Boston and Denver, CO, USA, 4Molecular, Cellular, and Developmental Biology, UCLA, Dana-Farber Cancer Institute, Boston, MA, USA Los Angeles, CA, USA Being able to generate functional β-cells from both human embryonic RATIONALE: The airway epithelium is in direct contact with the environ- (hESC) and induced pluripotent stem (iPSC) cells represents an important ment and constantly at risk for injury from toxins and infectious agents. milestone for the treatment of type I diabetes, not just because it could An efficient repair system is therefore needed to protect the host. Basal provide an unlimited source for β-cells, but also because it would provide cells have been found to repair the surface epithelium, but the potential a tool for drug discovery. Despite the recent success in differentiation of contribution of other stem cell populations to airway epithelial repair have hPSCs to the pancreatic lineages, the regulation of β-cell development and not yet been elucidated. METHODS: We used a model of severe hypoxic- maturation remains poorly defined. As a consequence, it is not yet possible ischemic injury and repair to identify cells that survive the injury and repair to efficiently and reproducibly generate fully functional β-cells in vitro and regenerate the airway epithelium. We found that airway submuco- and expansion of the current differentiation protocols to a broad range of sal gland duct cells, in addition to some basal cells, survived the severe human pluripotent stem cells is often hampered by inherent differences hypoxic-ischemic injury. We therefore hypothesized that submucosal gland between these stem cell lines. To address these issues, we investigated the duct cells may represent a stem/progenitor cell population and developed a role of specific signaling pathways at key stages of pancreatic differentia- method to isolate submucosal gland duct cells from the airway. We then used tion using five different hPSC lines and defined a protocol that can be used in vitro sphere and air-liquid interface models to compare the self-renewal broadly and efficiently to recapitulate the early stages of endocrine lineage and differentiation potential of submucosal gland duct cells to basal cells. We commitment in vitro. We focused on two important stages of pancreatic also developed a novel in vivo mouse model to examine the self-renewal and development: endoderm patterning and endocrine lineage commitment and differentiation capacity of submucosal gland duct cells as compared to basal on two major signaling pathways: TGFβ and WNT. We found that appropri- cells. In addition, we used lineage tracing of keratin(K)14-expressing submu- ate levels of WNT and BMP signaling are required for the correct pattern- cosal gland duct cells to identify the contribution of duct cells to the repair ing of the pancreatic epithelium. Low level of WNT signaling patterns the of the airway epithelium. RESULTS: We found that submucosal gland duct hPSC-derived endoderm to the pancreatic epithelium, while in response to cells were capable of forming spheres with cell types that expressed K5, K14, high concentration of WNT signaling hPSC-derived endoderm cells acquire TROP2, p63, K15, K8, and pIgR and produced mucins, as seen with Alcian a posterior/intestinal phenotype. Moreover, by using several hPSC lines, we Blue-Periodic Acid Schiff staining. In the in vivo model system, we found that uncovered a cell-line specific requirement for BMP inhibition prior to PDX1 submucosal gland duct cells, but not the basal cells, were capable of regen- induction. Treatment with Dorsomorphin, a BMP inhibitor, is essential for erating submucosal gland tubules and ducts. Lineage tracing demonstrated pancreatic patterning of H1, H9 and HES3 cell lines a finding that demon- that K14-expressing submucosal gland duct cells repaired the submucosal strates that endogenous signaling can compete with pro-differentiation fac- glands, submucosal gland ducts and the surface epithelium overlying the tors. Following PDX1 induction, endocrine lineage commitment is achieved submucosal glands. CONCLUSIONS: We have identified a novel multipotent by inhibition of TGFβ signaling. We clearly show that inhibition of SMAD2 stem/progenitor epithelial cell population of the airway. This submucosal phosphorylation leads to an increase in cell survival/proliferation, while inhi- gland duct epithelial stem/progenitor cell population is capable of regenerat- bition of SMAD1/5/8 phosphorylation is crucial for endocrine lineage com- ing the submucosal glands, submucosal gland ducts and the surface airway mitment. Optimal stage-specific manipulation of WNT and TGFβ signaling epithelium overlying the submucosal gland ducts. This is of importance to the pathways resulted in a striking 250-fold increase in the levels of insulin field of lung regeneration as determining the repairing cell populations of the expression and yielded populations containing up to 25% C-Peptide+ cells. airway epithelium could lead to the identification of novel therapeutic targets By using a reporter cell line where GFP cDNA is targeted to the INS locus and cell-based therapies for patients with airway diseases. we were able to sort the INS:GFP+ cells and follow their development in vivo. Transplantation experiments revealed that INS:GFP+ cells generated under these conditions gave rise to glucagon-expressing cells in vivo, sug- gesting that the endocrine cells generated in vitro represent an early stage of pancreatic development.

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Detailed Program and Abstracts – Friday, June 17

THE ENDOTHELIAL CELL NICHE COORDINATES GENERATION OF ANTERIOR FOREGUT AND ITS HEPATIC SPECIFICATION THROUGH DUAL DERIVATIVES FROM HUMAN PLURIPOTENT REPRESSION OF WNT AND NOTCH SIGNALING CELLS Han, Songyan1, Dziedzic, Noelle1, Gadue, Paul2, Keller, Gordon3, Snoeck, Hans-Willem E.1, Green, Michael D.2, Chen, Antonia2, Gouon-Evans, Valerie1 Nostro, Maria-Cristina3, d’Souza, Sunita L.2, Schaniel, Christoph2, 1Mount Sinai School of Medicine, New York, NY, USA, 2Children’s Hospital Lemischka, Ihor R.2, Gouon-Evans, Valerie2, Keller, Gordon3 of Philadelphia, Philadelphia, PA, USA, 3University Health Network, 1Dept. of Oncological Sciences and Black Family Stem Cell Institute, Toronto, ON, Canada Mount Sinai School of Medicine, New York, NY, USA, 2Mount Sinai 3 A complex cross-talk between the endoderm and the close microenviron- School of Medicine, New York, NY, USA, Division of Stem Cell and ment is an absolute requirement to orchestrate hepatic specification and Developmental Biology, McEwen Centre for Regenerative Medicine, expansion. Studies in the mouse have shown that the adjacent septum , Toronto, ON, USA transversum and cardiac mesoderm through secreted FGF and BMPs Directed differentiation of human embryonic stem (hES) cells and human instruct the ventral endoderm to become hepatic endoderm. Consecutively, induced pluripotent state (hiPS) cells captures in vivo developmental path- endothelial cells have been shown to promote expansion of the specified ways for specifying lineages in vitro, thus avoiding perturbation of the ge- hepatic endoderm. Using a mouse ES cell line expressing GFP from the nome with exogenous genetic material. Thus far, derivation of endodermal brachyury (Bry) locus and a truncated human CD4 cDNA from the Foxa2 lineages has focused predominantly on hepatocytes, pancreatic endocrine locus (GFP-Bry/CD4-Foxa2 ES cells), we have established conditions for the cells and intestine. The ability to differentiate pluripotent cells into anterior efficient and reproducible induction and hepatic specification of definitive foregut derivatives would expand their utility for cell-based therapy and ba- endoderm. In these studies, activin-A-induced progenitors for definitive en- sic research to tissues important for immune function, such as the thymus; doderm were isolated as a GFP-Bry+/CD4-Foxa2high/c-Kithigh population. We for metabolism, such as thyroid and parathyroid; and for respiratory func- showed that BMP4, in concert with bFGF and activin-A, is required for he- tion, such as trachea and lung. We find that dual inhibition of TGF-β and patic specification of the progenitor for endoderm. In that study, endothelial BMP signaling after specification of definitive endoderm from pluripotent cells were always seen surrounding the hepatic colonies, and their presence cells results in a highly enriched anterior foregut endoderm population that was associated with hepatic endoderm expansion. To investigate the role of is uniquely competent to subsequently be patterned along dorsoventral and endothelial cells in the growth, survival and specification of ES cell-derived anteroposterior axes. These findings provide a platform for the generation endoderm, we used a reporter ES cell line in which the human CD25 was of anterior foregut endoderm derivatives. targeted to the Foxa3 locus in GFP-Bry/CD4-Foxa2 ES cell line. This line allowed isolation of committed endoderm cells that co-express hCD4-Foxa2 and hCD25-Foxa3 and did not generate endothelial cells. We reconstituted an in vitro culture system in which isolated committed endoderm cells Plenary VI – Stem Cell Metabolism and Aging were co-cultured with endothelial cells in the context of cardiac mesoderm and septum transversum instruction, as BMP4 and bFGF were present in Friday, June 17, 4:00 – 6:00 p.m. the media. In this culture setting, we provide mechanistic evidence that endothelial cells function not only to promote hepatic endoderm expansion SOX17 EXPRESSION CONFERS SELF- but also hepatic specification of endoderm in part through regulating Wnt RENEWAL POTENTIAL AND FETAL STEM and Notch pathways. Activation of Wnt and Notch by chemical or genetic CELL CHARACTERISTICS UPON ADULT approaches increases endoderm cell numbers but inhibits hepatic specifica- tion, and conversely chemical inhibition of both these pathways enhances HEMATOPOIETIC PROGENITORS hepatic specification and reduces proliferation. We showed that the effects Morrison, Sean J., He, Shenghui, Kim, Injune, Lim, Megan S. of Wnt and Notch on endoderm expansion and hepatic specification were additive and dependent on endothelial cells. Using identical co-culture HHMI and University of Michigan Center for Stem Cell Biology, Ann Arbor, conditions, we defined a similar dependence of endoderm cells harvested MI, USA from E8.25 embryos on endothelial cells to support their growth and A key question concerns the mechanisms that determine temporal identity hepatic specification. High throughput analyses comparing the co-cultured in stem cells. Fetal hematopoietic stem cells (HSCs) differ from adult HSCs endoderm with the endoderm grown alone are ongoing and will further in terms of gene expression profile, surface marker expression, differentia- give insights into the cross-talk-dependant hepatic specification. tion, and self-renewal capacity. We previously showed that the transcrip- Together, these findings (1) confirm a conserved role of Wnt repression for tion factor Sox17 is expressed by fetal, but not adult HSCs, and is required mouse hepatic specification, (2) uncover a novel role for Notch repression for the maintenance of fetal and neonatal HSCs, but not adult HSCs (Cell in the hepatic fate decision, and (3) demonstrate that repression of Wnt 130:470). In the current study we show that ectopic expression of Sox17 in and Notch signaling in hepatic endoderm is controlled by the endothelial adult HSCs and transiently reconstituting multipotent progenitors was suf- cell niche. The approach used in this study reinforces the power of the ES ficient to confer increased self-renewal potential and the expression of fetal cell system combined with in vivo animal studies to dissect developmental HSC genes, including fetal HSC surface markers. Sox17 expression enabled processes, and to ultimately generate functional ES cell-derived hepatic cells transiently reconstituting adult progenitors to give long-term multilineage clinically relevant for liver cell therapy. reconstitution that resembled fetal hematopoiesis, including increased erythropoiesis, increased myelopoiesis, and decreased lymphopoiesis. Long- term ectopic expression of Sox17 eventually led to leukemogenesis. These data demonstrate that Sox17 is sufficient to confer fetal HSC characteristics upon primitive adult hematopoietic progenitors and is therefore a key deter- minant of fetal HSC identity.

81 ISSCR 9th Annual Meeting www.isscr.org

Detailed Program and Abstracts – Friday, June 17 and Saturday, June 18

MODIFYING REGENERATIVE POTENTIAL AND chemical screening, and co-culture assays in a search for robust protocols that enhance the self-renewal of muscle stem cells, promote their myogenic CELL FATE WITHIN MYOGENIC LINEAGE differentiation, and enable their efficient transplantation and engraftment. Conboy, Irina M., Conboy, Michael, Paliwal, Preeti, Silva, These ongoing studies point to discrete biochemical pathways, including Haroldo, Jeong, Jaemin, Yousef, Hanadie both intrinsic changes in gene expression, local interactions with non- myogenic cell types, and systemic regulators, which together determine University of California, Berkeley, Berkeley, CA, USA the effectiveness of satellite cell maintenance and myogenic commitment Skeletal muscle stem, satellite cells have been identified more than 40 years throughout life. Finally, our recent results in aged, obese, and calorie ago and have provided an excellent model to studies of aging and disease. restricted animals suggest that satellite cell function may be profoundly The normal progression of postnatal myogenesis is from a satellite cell to influenced by metabolic experience, and that alterations in metabolism can myoblast to myotube, and interestingly we show that the reversal of dif- directly modulate satellite cell activity endogenously and in transplantation ferentiated state from myotubes back to myoblasts and from myoblasts to settings. Importantly, this metabolic regulatory axis appears to be conserved muscle stem like cells can be experimentally orchestrated by small molecules for stem cells housed in other regenerative tissues, such as the blood, and in and chemically defined biomatrix. Such strategies work in vitro and in vivo. both mice and humans. Thus, these studies highlight novel mechanisms by Studies of myogenesis in young versus aged mice and humans also revealed which stem cell activity may be coordinated with the physiological demands that muscle satellite cells do not age as badly/quickly as their stem cell niche of the tissues these cells support, and suggest new strategies for therapeutic (myofibers and blood serum). Interestingly aged differentiated niches (local intervention to enhance endogenous repair activity or improve the efficacy tissue and systemic) inhibit their own dedicated stem cells, thus thwarting of transplantation-based cell replacement. the process of tissue maintenance and regeneration. The age-dependent regulation of muscle stem, satellite cells by their niches involves the interac- tive Notch, TGF-beta, Wnt and MAPK pathways. In addition to aging, the regenerative potential of satellite cells becomes inhibited when glucose Plenary VII – Therapeutic Approaches to Stem Cells metabolism is perturbed in a mouse model of type 1 diabetes, which works Supported by Pfizer Neusentis and our Canadian colleagues through myostatin signaling pathway. Very interestingly, human embry- onic stem cells produce secreted proteins of high therapeutic value, which Saturday, June 18, 9:00 – 11:30 a.m negate the inhibition by the aged niches, thus rejuvenating tissue regenera- tion. STIMULATION OF THE WNT-PGE2 PATHWAY TO IMPROVE HSC SELF-RENEWAL EPIGENETIC REGULATION OF HEMATOPOIETIC Zon, Leonard I. STEM CELLS Stem Cell Program and Hematology/Oncology, Children’s Hospital and Dana-Farber Cancer Institute, HHMI, Harvard Stem Cell Institute, Harvard Goodell, Margaret A. Medical School, Boston, MA, USA Baylor College of Medicine, Houston, TX, USA Hematopoietic stem cells are used clinically for the rescue of the immune Hematopoietic stem cells (HSCs) reside in the bone marrow and are defined system after high dose chemotherapy for leukemia or lymphoma. HSCs by their capacity for lifetime maintenance of the blood and bone marrow, have the ability to self-renew, although the mechanism of this process but the mechanisms that control differentiation vs. self-renewal are still remains to be understood at a molecular level. Definitive hematopoietic poorly understood. DNA methylation of cytosine-phosphate-guanine (CpG) stem cells are derived during embryogenesis and are influenced by distinct dinucleotides is a key epigenetic modification that influences tissue-specific signals in the niche of the embryonic aorta. We have demonstrated that the and / or context-specific gene expression. Dnmt1 is the principal mainte- wnt-PGE2 pathway can induce HSCs during embryogenesis. Adult marrow nance methyltranferase that re-establishes methylation marks during DNA treated with wnt agonists or dmPGE2 leads to increased engraftment in replication, and its loss leads to nearly immediate and complete loss of HSC competitive transplantation studies in mice and zebrafish. Human cord activity in vivo. However, the role of de novo DNA methylation has not blood is a source of HSCs, but few cells are obtained in a typical harvest. In been thoroughly explored. We have found that ablation of either Dnmt3a an effort to enhance stem cell engraftment of cord blood, a clinical trial has or Dnmt3b leads to expansion of HSC and loss of self-renewal activity, been undertaken to treat cord blood with dmPGE2, and evaluate engraft- demonstrating that DNA methylation is critical for stem cell differentiation ment over time. In an effort to understand the targets of the wnt pathway, in HSC, in parallel to its role in ES cells. The phenotypic as well as molecular we have undertaken Chip seq studies of TCF4. Surprisingly, TCF4 binds to impact of loss of de novo DNA methylation will be presented. cell-specific genes adjacent to the cell-specific transcriptional regulators. This provides a mechanism for how developmental signals such as wnt affect cell differentiation and self-renewal. We also examined the BMP MODULATORS OF STEM CELL REGENERATIVE signaling pathway that are also involved in hematopoietic induction during embryogenesis. Chip seq analysis demonstrated that SMAD1, similar to FUNCTION IN SKELETAL MUSCLE TCF4, binds to cell-specific genes. Our work suggests that signaling tran- Wagers, Amy scription factors function together with cell-specific factors to regulate the intrinsic hematopoietic program. These studies may help develop a clinical Harvard University and Joslin Diabetes Center, Boston, MA, USA therapy that could enhance engraftment of HSCs during transplantation. Skeletal muscle is a highly specialized tissue composed of non-dividing, multi-nucleated muscle fibers, as well as specialized muscle-forming stem cells (satellite cells) that remain associated with muscle fibers and are responsible for muscle growth and repair throughout life. Using direct cell sorting technologies, we have isolated satellite cells from muscle tissue and shown that they can be used to regenerate diseased muscle tissue. In addition, we find that dysfunction of these cells during aging contrib- utes to age-associated muscle disease. To identify molecular pathways that regulate satellite cell function, we combined transcriptional profiling,

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Detailed Program and Abstracts – Saturday, June 18

DRIVING HEART PROGENITOR FATE IN VIVO of the first clinical trial investigating the use of tissue engineered vascular grafts in humans. Next we will highlight our recent work investigating the WITH MODIFIED RNA cellular and molecular mechanisms underlying neovessel formation. Finally Chien, Kenneth R. we will demonstrate how these mechanistic studies are leading to discover- ies that are being applied to the rationale design of next generation tissue MGH Cardiovascular Research Center and the Dept of Stem Cell and engineered vascular grafts. Regenerative Biology, Harvard University, Boston, MA, USA A family of islet-1 and other multipotent heart progenitors are responsible for the diversification and expansion of distinct cardiac muscle, vascular LIMBAL STEM-CELL THERAPY AND LONG- smooth muscle, and endothelial cell lineages during murine and human cardiogenesis. Although a rare number of heart progenitors are found in the TERM CORNEAL REGENERATION post-natal heart, previous cell or molecular based approaches to activate De Luca, Michele1, Rama, Paolo2, Pellegrini, Graziella1 these or other endogenous cardiac progenitors following cardiac injury have 1Centre for Regenerative Medicine “Stefano Ferrari”, University of Modena had limited success. Toward this goal, we have identified VEGF-A as a key 2 switch in the human fetal heart that expands the vascular progenitor pool in and Reggio Emilia, Modena, Italy, Ophthalmology Unit, San Raffaele the family of Islet-1 human heart progenitors during ES cell cardiogenesis, Scientific Institute, Milan, Italy and have utilized modified RNA to transiently express the corresponding Epithelial stem cells are responsible for the continuous renewal and repair of protein in cardiac muscle in vitro and in vivo. In vitro, both neonatal and human stratified epithelia, are clonogenic and are known as holoclones. The adult mouse cardiomyocytes, as well as human fetal cardiomyocytes, can be stem cells of the human corneal epithelium are located in the limbus, the highly efficiently transfected (70-90%) with a transient, non-immunogenic, narrow zone between the cornea and the bulbar conjunctiva. Self-renewal modified mRNA (MOD RNA), with minimal toxicity or triggering of in- and proliferation of limbal stem cells are regulated by the ΔNp63 (α, β and nate immunity, and enhanced protein production vs non modified RNA. γ), C/EBPδ and Bmi1 transcription factors. In vivo, direct intramuscular injection of a luciferase reporter MOD RNA Ocular burns may destroy the limbus, causing limbal stem-cell deficiency in either cardiac or skeletal muscle results in a time and dose dependent (LSCD). In such cases, the cornea acquires an epithelium through the inva- expression detectable within 3 hours, persisting over several days, and sion of bulbar conjunctival cells. This process causes neovascularization, returning to negligible levels. In addition, injection of a Cre MOD-RNA chronic inflammation, and stromal scarring, leading to corneal opacity and into the hearts of Rosa26R-LacZ indicator mice revealed that the transfec- loss of vision. Allogeneic corneal transplantation, aimed at replacing the tion could drive expression in a wide region, well beyond the initial site of scarred corneal stroma and the inner endothelium, is not in itself a success- injection. Transfection of human or murine cardiomyocytes with hVEGF-A ful treatment. Although it temporarily removes the opacity, the conjunctival MOD RNA induced endothelial-like cells to form tube-like structures which cells will resurface the cornea. The only way to prevent this invasion is to were CD31+, VE-cadherin+ and vWF+, whereas most cardiac cells treated restore the limbus. The finding that human limbal cell cultures contain stem with vehicle were vimentin+. By injecting hVEGF and Cre MOD RNA into cells (detected as holoclones) led to the first therapeutic use of such cultures the Rosa26R-LacZ hearts after myocardial infarction, we have shown that in the regeneration of corneal epithelium. most of the cells in the injected area were positive for LacZ, and marked proliferation of a population of endothelial, smooth muscle, and cardiac Here we report long-term (up to 10 years) clinical results obtained in an myocytes, providing direct evidence of an in vivo regenerative response. homogeneous group of 112 patients presenting with corneal opacification Moreover, new proliferating vessels which are LacZ +were easily visual- and visual loss due to chemical burn-dependent LSCD (86.6% unilateral ized on the epicardial surface from the initial site of injection, documenting and 13.4% bilateral) and treated with autologous limbal stem cell cultures. that the transfected cells within the heart were adopting both a cardiac Clinical results were assessed by means of Kaplan-Meier, Kruskal-Wallis, myogenic and vasculogenic fate. In summary, MOD RNA is a new platform and univariate and multivariate logistic-regression analyses. to allow the rapid in vivo assay of known or novel paracrine protein factors Permanent restoration of a transparent, renewing corneal epithelium was that drive endogenous heart progenitor mobilization following heart injury. attained in 76.6% of eyes. The failures occurred within the first year. Further studies are warranted to evaluate whether this approach can be Restored eyes remained stable over time, with up to 10 years of follow- extended to other organ systems and whether hVEGF MOD RNA is a new up (mean, 2.91±1.99; median, 1.93). In post hoc analyses, success was therapeutic paradigm to achieve the recruitment and subsequent differen- associated with the percentage of holoclone-forming stem cells in culture. tiation of endogenous heart progenitors for cardiovascular regeneration. Cultures in which stem cells (detected as p63-bright cells) constituted more than 3% of the total number of clonogenic cells were associated with the permanent regeneration of a functional corneal epithelium in 80% of THE DEVELOPMENT AND TRANSLATION OF patients. In contrast, cultures in which such cells made up 3% or less of the THE TISSUE ENGINEERED VASCULAR GRAFT: total number of cells were associated with successful transplantation in only 10% of patients. Graft failure was also associated with the type of initial FROM THE BENCH TO THE BEDSIDE AND ocular damage and postoperative complications. BACK AGAIN We also show that cultures established from a single 1-2 mm limbal biopsy Breuer, Christopher K. offer an opportunity to treat virtually blind patients who have severe Yale University School of Medicine, New Haven, CT, USA bilateral loss of corneal epithelium, provided that a tiny part of the limbus is spared in one of the two eyes. Tissue engineering provides a method for creating tissue from its cellular components. Tissue can be created by seeding autologous cells onto a biodegradable three-dimensional scaffold. The scaffold provides sites for cell attachment and space for neotissue formation. Using tissue-engineering techniques, we have created the first man-made vascular graft with growth potential. This man-made vascular graft holds great promise for advancing the field of congenital heart surgery where the need for graft replacement due to somatic overgrowth is a significant source of morbidity and mortal- ity. In this presentation we will review the preclinical studies and results

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Detailed Program and Abstracts – Saturday, June 18

A CHEMICAL APPROACH TO CONTROLLING channel SCN5A is conserved between rodents and humans, providing an opportunity to compare stem cell derived cardiomyocytes with their primary CELL FATE counterparts. We demonstrated that the electrical phenotype of primary Ding, Sheng cardiomyocytes heterozygous for an SCN5A mutation is the same as car- diomycoytes from heterozygous mouse ESCs and iPSCs and from patients Gladstone Institute of Cardiovascular Disease, Dept of Pharmaceutical with the same mutation, indirectly validating the value of hiPSC and hESC. Chemistry, University of California San Francisco, La Jolla, CA, USA However, to encourage the development and use of alternative cardio- Recent advances in stem cell biology may make possible new approaches myocyte sources based on hESC or hiPSC by the pharmaceutical industry, for the treatment of a number of diseases. A better understanding of mo- it is essential that cardiomyocyte production is (i) defined (ii) reproducible lecular mechanisms that control stem cell fate as well as an improved ability and (iii) scalable to high throughput formats. We are using a hESC line with to manipulate them are required. Toward these goals, we have developed GFP targeted to the Nkx2.5 locus for this purpose. Nkx2.5 is a transcription and implemented novel high throughput phenotypic screens of chemical factor expressed in cardiac progenitor cells with the capacity to form car- libraries to identify and further characterize small molecules that can control diomyocytes, endothelial cells and vascular smooth muscle cells depending stem cell fate in various systems. This talk will provide latest examples of on the signaling pathways activated and suppressed. Using combinations discovery efforts in my lab that have advanced our ability and understand- of growth factors and signaling pathway inhibitors we are able to gener- ing toward controlling stem cell fate, including self-renewal, survival, dif- ate populations of hESC derivatives containing up to 80% cardiomyocytes ferentiation and reprogramming of pluripotent stem cells. without selection in completely defined media. This “best in class” protocol is proving effective in hiPSCs, helping address one of the present hurdles to full implementation of these cells in this area. TITLE TBD Sabine, Charles NONVIRAL CORD BLOOD-DERIVED Patient Advocate,UK IPSC POSSESS AUGMENTED CAPACITIES ABSTRACT NOT AVAILABLE AT TIME OF PRINTING FOR GENERATING FUNCTIONAL AND ENGRAFTABLE ANGIOBLASTS IN A RETINAL Concurrent Session III Track A – Cell Therapy ISCHEMIA MODEL Park, Tea Soon1, Bhutto, Imran2, Miller, Diana3, Feldman, Supported by F. Hoffman-La Roche Ltd. Ricardo3, Lutty, Gerard2, Zambidis, Elias1 Saturday, June 18, 1:30 – 3:15 p.m. 1Institute for Cell Engineering, Johns Hopkins School of Medicine, Baltimore, MD, USA, 2Department of Ophthalmology, Johns Hopkins CARDIOMYOCYTES FROM PLURIPOTENT STEM School of Medicine, Baltimore, MD, USA, 3University of Maryland, CELLS IN GENETIC CARDIAC DISEASE AND Baltimore, MD, USA DRUG SAFETY PHARMACOLOGY Retinal ischemic diseases are caused by dysfunction of endothelial/peri- vascular cells and result in deterioration of ophthalmic functions. Although 1 1 1 Mummery, Christine L. , Davis, Richard , Casini, Simona , cell-based therapies using angioblasts or endothelial progenitor cells (EPC) Tertoolen, Leon1, Passier, Robert1, Braam, Stefan1, Freund, have the potential to promote vascular repair of ischemic retina, the source Christian1, van den Berg, Cathelijne1, Dambrot, Cheryl1, Atsma, of progenitors is limited. Nonviral, non-integrated human induced pluripo- Douwe1, Ward, Dorien1, Bezzina, Connie2, Wilde, Arthur2, Elliott, tent stem cells (hiPSC) may provide clinically safe EPC for the treatment of David3, Ng, Elizabeth3, Elefanty, Andrew3, Stanley, Ed3 retinal ischemic diseases. Furthermore, the use of hiPSC derived from cord blood progenitors has the potential to generate more robust amounts of 1Leiden University Medical Center, Leiden, Netherlands, 2Academic functional EPC due to epigenetic memory of its original hemato-endothelial Medical Centre, Amsterdam, Netherlands, 3Monash University, Melbourne, lineage. In this study, we generated EPC from human embryonic stem cells Australia (hESC), fibroblast-derived iPSC, and nonviral CD34+ CB-iPSC and exam- Cardiomyocytes derived from both embryonic and induced pluripotent stem ined the comparative functional and engrafting potential of EPC in a mouse cells show many functional properties of their primary cell counterparts retinal ischemia/reperfusion injury model. even though relatively immature. These may include their electrophysi- HESC (H9), fibroblast-iPSC (IMR90-1), and CB-iPSC were differentiated via ological properties such as action potentials and transmembrane current human embryoid bodies (hEB), and further differentiated into vascular lin- densities. Disease mutations can affect these functional phenotypes: ion eages on fibronectin-coated culture plates in endothelial growth medium-2 channel mutations for example cause alterations in action potentials of (EGM2). Thereafter, the monolayer included cells expressing hematopoietic, cardiomyocytes that are thought to reflect channelopathies seen in cardiac vascular, and endothelial markers (CD31, CD34, CD146, α-SMA, KDR/ patients. These functional assays in principle provide platforms for identify- VEGF-R2, CD133, and BB9/CD143). The presence of putative EPC was ing compounds or drugs that affect normal function of these cells as well evaluated from these cells using endothelial (CD31) and pericytic (CD146) as compounds or interventions that return functional behaviour to normal markers. Four purified populations based on expression of CD31 and when the cells carry disease mutations. Examples will be given of how CD146 were thoroughly analyzed for endothelial function (EPC colony cardiac derivatives of hESC respond to drugs in clinical use. These include assay, Matrigel microvessel tubular formation, Dil-acetyl-LDL uptake, and cardiac drugs as well as non-cardiac drugs removed from clinical practice flow cytometry analysis). Based on these analyses, CD31+/CD146+ popula- because of adverse effects on the heart. That 30% of drugs are withdrawn tion were determined to represent a highly functional angioblast-containing late in development demonstrates that current test systems based on population, and found to be more robustly generated from CB-iPSC. We either single ion channels expressed in tumor cell lines or rodent or canine next differentiated CD31+/CD146+ populations from hESC and iPSC lines cardiomyocytes do not detect risk adequately, reducing their value in drug that were lentivirally-transduced with a luciferase reporter, and transplanted discovery. This is in part because ion channels are utilized differently in them into an adult murine retinal ischemia/reperfusion injury. Human cells humans than in most animal species and in part because correct ion channel were easily distinguished from mouse tissue via luciferase immunostain- function requires the complex environment of a cardiomycoyte. Potassium ing. To visualize the vascular injury in flat mount retinas, we stained viable channel use is a a specific example. Exceptionally, the activity of the sodium 84 www.isscr.org Final Program

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blood vessels with rat anti-mouse CD31 and all retinal capillaries (viable and and HUCPVCs, resp. vs 50±8% with medium; p<0.05) but not in the BM. acellular) with anti-collagen IV. Three-, 7-, and 14-day post-transplantation Moreover, the infarct site exhibited decreased expression of IL-1β and IL-6, results revealed that CD31+/CD146+ human cells engrafted into the dam- increased IL-10 and reduced cardiomyocyte apoptosis, without changes in aged mouse retinal blood vessels, while roaming human cells were observed angiogenesis. These observations were associated with enhanced cardiac in the vitreous body of non-injured control mouse eyes. Interestingly, CB- function (fractional shortening) at 2 and 4 weeks after infusion but were iPSC were observed more robustly at longer transplantation time periods. similar to medium controls 16 weeks after MI. In vitro studies showed that These data indicated that CD31+/CD146+ cells were highly responsive to co-culture of BM-MSCs or its conditioned medium directly enhanced the the signals from injured cells, and actively migrated, and incorporated to the frequency of CD206+ M2 monocytes/macrophages and expression of damaged retinal vasculature. IL-10, TGF-β and arginase-1. The frequency of CD206 + cells was partially reduced by a human specific IL-10 antibody. In summary, we have successfully derived a functional and engraftable EPC population from CB-iPSC that possessed enhanced survival and integration Our data infer a new mechanism for MSC-mediated enhancement of into ischemic mouse eyes compared to other sources. The use of CB-iPSC cardiac function via an IL-10 mediated switch from pro-inflammatory to is more clinically applicable since this somatic source carries relatively few anti-inflammatory macrophages infiltrating the infarct site. We suggest that somatic mutations, and could be used to create an HLA-defined stem cell additional pre-clinical studies augmenting the anti-inflammatory effects of bank via worldwide networks of existing blood bank repositories. Thus, MSCs in cardiac regeneration are warranted. these studies provide a preclinical model for evaluating the potential of nonviral CB-iPSC for generating transplantable EPC for treating vascular degenerative disorders. THE PARACRINE MECHANISM OF ANTI- FIBROTIC AND ANTI-INFLAMMATORY EFFECTS MESENCHYMAL STROMAL CELLS MEDIATE OF HUMAN PERIVASCULAR STEM CELLS THE SWITCH TO ALTERNATIVELY ACTIVATED Chen, Chien-Wen1, Proto, Jonathan2, Gao, Xueqin3, Okada, MONOCYTES/MACROPHAGES AFTER ACUTE Masaho3, Corselli, Mirko4, Crisan, Mihaela3, Tobita, Kimimasa5, 4 3 MYOCARDIAL INFARCTION Peault, Bruno , Huard, Johnny 1Bioengineering, University of Pittsburgh, Pittsburgh, PA, USA, 2Pathology, Dayan, Victor1, Yannarelli, Gustavo2, Billia, Filio3, Filomeno, Paola2, University of Pittsburgh, Pittsburgh, PA, USA, 3Orthopedic Surgery, Wang, Xing-Hua2, Davies, John E.4, Keating, Armand2 University of Pittsburgh, Pittsburgh, PA, USA, 4David Geffen School of 1University of the Republic, Montevideo, Uruguay, 2University Health Medicine, University of California at Los Angeles, Los Angeles, CA, USA, 5 Network, Toronto, ON, Canada, 3Division of Cardiology, Campbell Family Developmental Biology, University of Pittsburgh, Pittsburgh, PA, USA Institute for Breast Cancer Research, University Health Network, Toronto, Perivascular cells, particularly pericytes, purified from multiple human or- 4 ON, Canada, Institute of Biomaterials and Biomedical Engineering, gans have been shown to harbor stem cells endowed with robust mesoder- University of Toronto, Toronto, ON, Canada mal developmental potentials. Transplantation of multipotent perivascular After acute myocardial infarction (MI), pro-inflammatory M1 monocytes/ cells was shown to repair and regenerate damaged/dystrophic skeletal macrophages (Ly6Ghi+ and expressing elevated IL-1β, IL-6 levels) and muscles in vivo as well as improve cardiac function post-infarction. Previ- anti-inflammatory M2 monocytes/macrophages (CD206+ and expressing ously we demonstrated that PSCs support the formation of microvascular elevated IL-10, TGF-β and arginase-1 levels) infiltrate ischemic myocardium. structures in vitro and promote angiogenesis in vivo after acute myocardial Given the established anti-inflammatory properties of mesenchymal stromal infarction (AMI). In the current study, we investigated the hypothesis that cells (MSCs), we wished to determine their effect on the subtype of inflam- transplantation of human perivascular stem cells (PSCs) results in anti- - - - + matory cells infiltrating ischemic cardiac tissue, hence on cardiac function. fibrotic and anti-inflammatory effects. Purified CD34 /45 /56 /146 PSCs 5 We employed 2 types of MSCs, human bone marrow-derived (BM) MSCs were injected into ischemic myocardium of immunodeficient mice (3.0×10 and human umbilical cord perivascular cells (HUCPVCs) in an experimen- cells). Using Masson’s trichrome staining, we estimated the anti-fibrotic tal acute MI model with immune deficient NOD/SCID gamma null mice effect of PSCs in vivo by the extent of myocardial fibrosis at 2 weeks post- (n=42). BM-MSCs (2x106 cells) (n=14), HUCPVC (2x106 cells) (n=14) or infarction. A significant 45.3% reduction of the scar area in PSC-injected medium (n=14) were infused i.v. 48 hours after induction of MI and as- hearts was observed when comparing to saline-injected controls (P≤0.001). sayed 72 hours later (n=6 per group) for monocytes/macrophages by flow Measurement of LV wall thickness at the center of the infarct indicated cytometry and immunohistochemistry in bone marrow, peripheral blood a 26.3% increase in the PSC group (0.255 ± 0.026 mm) versus the PBS and cardiac tissue. Expression of pro-inflammatory and anti-inflammatory group (0.202 ± 0.040 mm), suggesting the potential of PSCs to ameliorate cytokines, chemokines, adhesion factors and apoptosis was evaluated in the cardiac remodeling. Consequently we proposed a role of donor cell-derived MI area. Presence of human DNA was also assessed in the heart, lung, liver, matrix metalloproteinases (MMPs) and a paracrine mechanism in diminu- BM and spleen tissue. Echocardiographic evaluation of fractional shortening tion of fibrosis. To illustrate MMP levels in PSCs specifically under the was done at baseline (before the MI), 2, 4 and 16 weeks after MI. hypoxic microenvironment, cells were cultured in vitro under 2.5% oxygen NOD/SCID gamma null bone marrow-derived macrophages were co-cul- for 24 hours. Real-time quantitative PCR analysis showed a sustained high tured with BM-MSCs or with BM-MSC-conditioned medium. Macrophages expression of MMP-2 but not MMP-9 by PSCs under hypoxia. MTS assay were characterized after 3 days by flow cytometry and quantitative PCR. revealed significantly diminished cell growth of primary murine fibroblasts cultured with hypoxic PSC-conditioned media when comparing to cells We showed that in the presence of BM-MSCs and HUCPVCs, over- cultured with normoxic PSC-conditioned media (P=0.015), implicating all monocyte/macrophage levels were reduced in BM (31.2±1.7% and paracrine reduction of fibroblast proliferation by PSCs under hypoxia. 31.2±1.8%, respectively vs 38.5±1.8% with medium; p<0.05); in the The anti-inflammatory effect of PSCs was first revealed by H&E staining circulation (7.3±0.9% and 27±1.6%, respectively vs 36.6±0.6% with showing decreased infiltration of dark-nucleated inflammatory cells within medium; p<0.001) and in cardiac tissue (0.57±0.06% with BM-MSCs the infracted region of PSC-injected hearts. The infiltration of host CD68+ vs 1.65±0.65% with medium; p<0.05). The proportion of M2 macro- monocytes/macrophages was significantly inhibited within the infarct in phages was significantly increased in the circulation (21.5±1.3% and PSC-treated hearts when comparing to saline-injected hearts (P<0.001), 15.7±1.2% with BM-MSCs and HUCPVCs respectively, vs 4.9±2.5% with suggesting that reduction of inflammation contributed to cardiac restora- medium; p<0.05) and heart (77.2±2.2% and 81.3±2% with BM-MSCs tion. Moreover, when cultured with hypoxic PSC-conditioned media in vi-

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tro, the proliferation of murine monocytes/macrophages were significantly MECHANISTIC INSIGHTS INTO DEFINED- inhibited (P<0.001), providing further evidence of the immunomodulation by PSCs. To illustrate the potential paracrine mediators utilized by PSCs in FACTOR REPROGRAMMING USING SMALL ischemic tissue repair, we examined the expression of immunoregulatory MOLECULES

genes in PSCs cultured under hypoxic microenvironment (2.5% oxygen) in 1 1 1 2 vitro. Semi-quantitative RT-PCR analysis revealed sustained high expres- Ichida, Justin K. , Son, Esther Y. , Singh, Sean , Wainger, Brian , 1 1 1 3 sion of immunoregulatory genes in both normoxic- and hypoxic-cultured Blanchard, Joel , Lam, Kelvin , Chung, Julia , Toma, Jeremy , 3 2 1 1 PSCs, including IL-6, LIF, COX-2, HMOX-1, with no significant difference Rafuse, Victor , Woolf, Clifford , Meissner, Alexander , Rubin, Lee , (P>0.05). The expression of MCP-1, however, was notably repressed in Eggan, Kevin1 hypoxic-cultured PSCs (P=0.027). No expression of IL-10 and iNOS was 1Stem Cell and Regenerative Biology, Harvard Stem Cell Institute, detected in both normoxic- and hypoxic-cultured PSCs. Overall, our data Cambridge, MA, USA, 2Harvard University, Boston, MA, USA, 3Dalhousie suggest a role of PSC-mediated anti-scarring and anti-inflammatory effects University, Halifax, NS, Canada through paracrine mechanism in the ischemic myocardial repair. The combined activity of Oct4, Klf4, and Sox2 can reprogram adult cells into induced pluripotent stem cells (iPSCs). However, the potential of using defined-factor reprogramming to direct the differentiation of stem cells or STEMMING VISION LOSS USING STEM CELLS - directly generate other cell types is restricted because the molecular mecha- SEEING IS BELIEVING nisms governing reprogramming are poorly understood. These limitations could be overcome if biological pathways that mediate reprogramming Coffey, Peter could be identified and controlled. To this end, we performed small mol- Institute of Ophthalmology, University College London, London, United ecule screens in two defined-factor reprogramming systems; the conversion Kingdom of fibroblasts and keratinocytes to iPSCs and the conversion of fibroblasts to The London Project to Cure Blindness was launched at the UCL Institute motor neurons, to identify biological pathways that modulate reprogram- of Ophthalmology in June 2007, and aims to make the most of human ming in a general, starting cell-, or target cell-specific manner. embryonic stem cells to prevent blindness and restore sight in patients with We recently identified a cohort of 7 transcription factors that converts Age-related Macular Degeneration (AMD) by 2012. Our goal is to replace fibroblasts into bona fideHb9+ spinal motor neurons (MNs), the cells cells essential for “seeing” lost by disease at the back of the eye. We aim lost in Amyotrophic Lateral Sclerosis. To identify chemicals that modulate to repair and regenerate the aged diseased eye using human embryonic reprogramming, we transduced Oct4::GFP mouse fibroblasts or keratino- stem cells which have been transformed into the cells affected in AMD: the cytes with the iPSC factors and Hb9::GFP mouse fibroblasts with the 7 MN support cells for the photoreceptors (retinal pigment epithelium) and the factors in parallel. After treatment with an 800-member chemical library for photoreceptors. The cells will be surgically implanted into a clinical popula- 2 weeks, we scored wells with increased numbers of Oct4::GFP+ iPSC colo- tion of AMD patients. nies or Hb9::GFP+ MNs compared to controls and validated hit compounds by repeating the assay. Several molecules increased the conversion of fibroblasts to both iPSCs Concurrent Session III Track B – and MNs. Several Tgf-β inhibitors or vitamin C increased the number of iPSCs and motor neurons by 3-10-fold. Early chemical treatments before Small Molecule Approach to Stem Cells transduction were most effective, indicating they can act on the starting fibroblasts. In contrast, Tgf-β inhibition did not improve repro- Supported by Centre for Commercialization of gramming, indicating this effect is fibroblast-specific. These results suggest Regenerative Medicine that inhibition of Tgf-β signaling or vitamin C treatment may increase the plasticity of fibroblasts in defined-factor reprogramming. Saturday, June 18, 1:30 – 3:15 p.m Histone deacetylase (HDAC) inhibition also enhanced both iPSC and MN TOWARDS IDENTIFYING FACTORS THAT CAN generation. However, the efficacy of particular HDAC inhibitors depended heavily on the target cell type. Valproic acid increased iPSC generation by AID IN TISSUE REGENERATION 10-fold but did not affect MN induction. Conversely, trichostatin A was the Bouwmeester, Tewis, Ruffner, Heinz, Salathe, Adrian, Bouchez, most effective HDAC inhibitor for MN induction but only modestly affected iPSC formation. These results indicate that chromatin structure restricts Laure, Mueller, Matthias, Kinzel, Bernd, Latario, Brian, Lacoste, reprogramming and that the target cell type dictates the efficacy of HDAC Arnaud, Labow, Mark, Schebesta, Michael, Serluca, Fabrizio, Curtis, inhibitors. Daniel Notch inhibitors specifically affected only one type of starting cell. DAPT Novartis Institutes for Biomedical Research, Novartis Pharma AG, Basel, increased the conversion of keratinocytes to iPSCs by 5-10 fold, but had no Switzerland effect on fibroblasts. We found that DAPT acts by enriching for keratino- Stem cells hold great promise for regenerative therapy in a number of cyte progenitors and increasing Wnt signaling. degenerative conditions, such as chemotherapy- and radiation-induced Lastly, we identified chemicals that enhanced conversion specifically to mucositis of the gastro-intestinal tract, neurological disorders and cancer- one target cell state. Treatment with the GSK3β/Mek inhibitor cocktail 2i related muscle wasting. The pursuit for regenerative factors (low molecular enhanced the generation of iPSCs but did not increase MN induction. Thus, weight (LMW) compounds and/or secreted biological factors) that can 2i promotes conversion specifically to the iPSC state, which may reflect the activate/control stem cells under such patho-physiological conditions offers ability of Wnt signaling and Mek inhibition to maintain embryonic stem cell potential therapeutic benefit, but remains challenging. The identification of self-renewal. bona fide stem cell markers has been an important milestone in the study of tissue stem cell biology and has enabled screening for modulators of stem Together these results indicate that while chromatin structure restricts repro- cell number/activity. I will discuss some of approaches we use to screen gramming, inhibition of Tgf-β signaling or vitamin C treatment increases for novel low molecular weight and biological factors that have intrinsic the plasticity of fibroblasts, and controlling signaling pathways that govern regenerative activity. the maintenance of the target cell state can greatly facilitate defined-factor reprogramming. 86 www.isscr.org Final Program

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SOLUBLE FACTORS SPECIFY THE GENERATION Using our optimized culture conditions in 384-well-plates and automated qRT-PCR protocols, we tested the impact of chemical compound libraries OF HEMATOPOIETIC PROGENITORS WITH (>7000 compounds, FDA-approved drugs and natural products) on WT MULTILINEAGE ENGRAFTMENT POTENTIAL and MU IKBKAP expression in patient-specific and symptom-relevant NCs. IN ADULT RECIPIENT MICE FROM MOUSE Expression levels were normalized to housekeeping gene and all assays were performed in three biological repeats per compound. We identified EMBRYONIC STEM CELLS a total of 41 primary hits that were subjected to various validation studies, Heffner, Garrett C., Wenzel, Pamela, Daley, George Q. cytotoxicity assays and dose-response studies (12 concentrations). A total of 8 candidate compounds were selected for further detailed analysis, Children’s Hospital Boston, Boston, MA, USA and several of those compounds showed evidence of rescue of molecular The efficient generation of engraftable hematopoietic progenitor cells from defected such as mis-regulation of autonomic gene expression (ASCL1 and an in vitro culture system would represent a clinical advance in the treat- SCG10 phenotypes). Currently, we are i) testing the potential of those com- ment of patients with hematopoietic disorders. Our approach has involved pounds to rescue additional disease phenotypes (i.e. migration defects), ii) the well-established culture paradigm whereby embryonic stem cells (ESC) applying them to NCs derived from other FD-iPSC lines, iii) finding effects are differentiated as embryoid bodies (EBs). After six days of culture as EBs, on other non-NC lineage (such as hematopoietic, endothelial, endodermal cells were exposed to a panel representing over 1500 individual combina- and mesenchymal cells) and iv) understanding their intracellular actions tions of cytokines, small molecules, and soluble factors in 96-well dish through unbiased transcriptome analysis. format. After 24 hours of culture, cells were aggregated into pools of ~50 Our drug discovery effort using symptom-relevant and patient-specific cells combinations, and each pool was transplanted via tail vein into lethally is a proof of concept demonstration for HTS with disease-specific hiPSC irradiated adult Rag2-/- gammaC -/- mice, and engraftment was monitored lines. We hope that our proposed paradigm (specification/isolation of over twenty weeks via analysis of peripheral blood at staged timepoints. symptom-relevant cell type, disease modeling and HTS for drug discovery) While most pools yielded zero engraftment at any timepoint, we identified will prove suitable for other iPSC based disease models and lead to novel a small number of pools that yielded positive myeloid engraftment that per- treatments for many currently intractable diseases. In conclusion, our stud- sisted throughout the twenty-week analysis window. We also identified one ies illustrate the potential of using patient-specific hiPSCs for HTS-based pool that yielded short-term myeloid and persistent lymphoid engraftment drug discovery. throughout the analysis window. We anticipate that these findings and our experimental approach will serve as a model for high-throughput screening of soluble factors for specification of pluripotent stem cells into functional stem cells of many adult tissues for future clinical applications of regenera- STEM CELL THERAPEUTICS: WHEN WILL tive medicine. PROMISE BECOME PROFITS? Ezekowitz, Alan DISCOVERY OF POTENTIAL THERAPEUTIC Former SVP Merck and Co, USA COMPOUNDS FOR FAMILIAL DYSAUTONOMIA Stem cell therapies offer potentially disruptive ways of treating a variety of inflammatory and degenerative diseases. I will review some of these USING PATIENT-SPECIFIC AND SYMPTOM- advances and provide an analysis of what factors influence the commercial RELEVANT IPSC DERIVED NEURAL CREST value of these advances to biotech and major pharmaceutical companies. PRECURSORS Lee, Gabsang, Ramirez, Christina, Kim, Hyesoo, Djaballah, Hakim, Studer, Lorenz Concurrent Session III Track C – Genomic Integrity Memorial Sloan-Kettering Cancer Center, New York, NY, USA Saturday, June 18, 1:30 – 3:15 p.m. Patient-specific human induced pluripotent stem cells (hiPSCs) offer un- precedented promise in disease modeling and drug discovery by providing TELOMERE DYNAMICS AT THE INNER CELL unlimited number of symptom-relevant cells. Taking advantage of our novel MASS AND EMBRYONIC STEM (ES) CELLS methodology of direct differentiation and prospective isolation of neural crest (NC) from human pluripotent stem cells, we successfully modeled INDICATE A LINK BETWEEN TELOMERE Familial Dysautonomia (FD) with hiPSC technology. The results from these BIOLOGY AND PLURIPOTENCY studies offered previously unknown clues for disease pathogenesis and allowed us to validate several candidate drugs. To extend our approach Blasco, Maria A. beyond the validation of a few candidate compounds, we present here our Spanish National Cancer Research Centre (CNIO), Madrid, Spain data on the first high-throughput screening (HTS) drug discovery effort Murine embryonic stem (ES) cells have unusually long telomeres, much lon- performed in FD-iPSC derived symptom-relevant NC precursors ger than those in embryonic tissues. Here we address whether hyper-long FD-iPSCs differentiated into NC using our standard MS-5 differentiation telomeres are a natural property of pluripotent stem cells, such as those protocol and purified by flow cytometry using HNK1 (CD57) antibody. present at the blastocyst inner cell mass (ICM), or whether is a characteristic Enriched NC population was further expanded for 2 weeks yielding > 0.5 x acquired by the in vitro expansion of ES cells. We find that ICM cells under- 109 cells that were could be cryo-preserved prior to performing HTS assay. go telomere elongation during the in vitro derivation of ES-cell lines. This To establish culture condition in 384-well-plates, we optimized seeding is concomitant with a decrease in heterochromatic marks at telomeres. We density, proliferation rate and coating reagents. Furthermore, by using also found increased levels of the TRF1 telomere capping protein in cultured commercially available kits, automated protocols for RNA extraction and ICM cells before telomere elongation occurs, coinciding with expression qRT-PCR were developed to measure level of wild type (WT) and mutant of pluripotency markers. These results suggest that high TRF1 levels are (MU) IKBKAP expression. We selected qRT-PCR as the primary HTS readout associated to pluripotency, most likely to ensure proficient capping of the given that reduction in transcriptional levels of wild-type IKBKAP is closely newly synthesized telomeres. These results highlight a previously unnoticed correlated with FD disease symptoms. difference between ICM cells at the blastocyst and ES cells, and suggest

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that abnormally long telomeres in ES cells are likely to result from continu- that the selection pressures that stem cells experience in-vitro resemble the ous telomere lengthening of proliferating ICM cells locked at an epigenetic ones that lead to tumor formation in-vivo. As chromosomal aberrations in state associated to pluripotency. pluripotent stem cells have been suggested to increase their tumorigenicity, such aberrations are expected to affect the safety of cultured multipotent stem cells as well. FUNCTIONAL CONSEQUENCES OF * The first part of this research was recently published in Cell Stem Cell SOMATIC MUTATIONS IN HUMAN INDUCED (October 2010). PLURIPOTENT STEM CELLS Gore, Athurva1, Ruiz, Sergio2, Li, Zhe1, Fung, Ho-Lim1, Belmonte, GENOME-WIDE FUNCTIONAL SCREEN FOR Juan Carlos Izpisua2, Zhang, Kun1 REGULATORS OF DNA DAMAGE RESPONSE IN 1University of California - San Diego, La Jolla, CA, USA, 2Salk Institute for Biological Studies, La Jolla, CA, USA HUMAN HEMATOPOIETIC STEM CELLS 1 1 1 Defined transcription factors can induce epigenetic reprogramming of adult Milyavsky, Michael , van Delft, Mark F. , Gan, Olga I. , Buzina, 2 3 2 3 mammalian cells into induced pluripotent stem (iPS) cells. Recent stud- Alla , Kalatskaya, Irina , Ketela, Troy , Stein, Lincoln , Moffat, ies revealed that human iPS lines acquire chromosomal rearrangements Jason2, Dick, John E.1 after long-term culture and single nucleotide point mutations enriched in 1Ontario Institute for Cancer Research, University Health Network, cancer-related genes after short-term culture. However, the exact source of Toronto, ON, Canada, 2Department of Molecular Genetics, University of reprogramming-associated mutations remains unclear. It is also unknown Toronto, Toronto, ON, Canada, 3Ontario Institute for Cancer Research, if mutations are functional “drivers” for the reprogramming process or if Informatics and Bio-Computing, Toronto, ON, Canada they are “passengers” acquired due to clonal selection. Here we show that iPS cell lines acquired point mutations regardless of donor age, progeni- Hematopoietic stem cells (HSC), by virtue of their longevity, experience tor cell type, or amount of passaging, implying that multiple mechanisms many genotoxic insults over lifetime. As such, HSC must protect genome were responsible for reprogramming-associated mutations. We additionally integrity, otherwise accumulation of genetic aberrations may lead to characterized the mutational load present after embryonic stem cell differ- malignant transformation or bone marrow failure. We recently reported entiation, implying that other in vitro stem cell techniques could compro- that human cord blood-derived HSC safeguard against this outcome by mise genomic integrity. We functionally characterized the effects of several being exquisitely sensitive to DNA damage-induced apoptosis. Following reprogramming-associated mutations in key cancer and pluripotency genes. exposure to ionizing radiation human HSC exhibit a persistent DNA damage Our results emphasize the need for further rigorous work on mutational response and greater p53-dependent apoptosis as compared with lineage load during reprogramming and differentiation. Extensive genetic screen- committed progenitors. ing should become a standard procedure to ensure stem cell safety before Here, we report studies aimed to further characterize the molecular basis clinical use. of this observation. Using highly purified fractions of HSC and progenitors, we performed global gene expression profiling to define the transcriptional response of each fraction to irradiation. We also carried out a genome-wide GENE EXPRESSION PATTERNS REVEAL loss-of-function genetic screen using a library of 80,000 lentiviral shRNA vectors targeting >16,000 human genes in a cord blood-derived immortal- NOVEL TISSUE-SPECIFIC CHROMOSOMAL ized cells, whose DNA damage response resembles that of HSC. Among ABERRATIONS IN HUMAN PLURIPOTENT AND the hits that mediate radiation resistance we identified known regulators of MULTIPOTENT STEM CELLS DNA damage response (e.g. p53), as well as numerous genes that were not previously connected to genotoxic stress response. Ben-David, Uri, Mayshar, Yoav, Benvenisty, Nissim An integrated analysis of these studies revealed a number of molecular The Hebrew University, Jerusalem, Israel networks, whose role in HSC DNA damage response is currently under Chromosomal aberrations are a potent driving force for cancer development investigation. and tumor progression. Stem cells are routinely expanded in-vitro, subject- ing them to selection pressures that might affect their genomic stability. We have recently developed a method to detect chromosomal aberrations in HAIR FOLLICLE STEM CELLS USE DIFFERENT stem cells based on their gene expression patterns. Using this methodology, MECHANISMS TO MEDIATE GENOME we analyzed over 200 samples of human pluripotent stem cells from over 25 studies. Our analysis reveals that chromosomal aberrations in human MAINTENANCE DEPENDING ON THEIR STAGES induced pluripotent stem cells occur frequently, and can originate from OF ONTOGENY three different sources: aberrations of somatic origin, aberrations that occur during or immediately after the reprogramming, and aberrations acquired in Blanpain, Cédric, Candi, Aurélie, Sotiropoulou, Panagiotta culture (only the latter is common in human embryonic stem cells). We next IRIBHM, Université Libre de Bruxelles, Bruxelles, Belgium analyzed over 400 samples of neural, mesenchymal and hematopoietic stem Adult stem cells (SCs) are at high risk of accumulating deleterious mutations cells from over 50 studies. Our analysis reveals chromosomal aberrations in because they reside and self-renew in adult tissues for extended periods of over 5% of the cells lines, and shows that only few passages are needed for time. Little is known about how adult SCs sense and respond to DNA dam- these aberrations to take over the culture. We identify novel aberrations in age within their natural niche. Using mouse epidermis as a model, we de- different types of multipotent stem cells, some of which recur in indepen- fined the functional consequences and the molecular mechanisms by which dent studies. Moreover, these aberrations are tissue-specific, as each type adult SCs respond to DNA damage. We found that multipotent hair follicle of stem cells is prone to acquire a unique set of chromosomal aberrations. bulge SCs exhibit increased resistance to DNA damage-induced cell death Interestingly, the same aberrations also arise in the respective tissue-specific by the higher expression of the anti-apoptotic gene Bcl2 and the transient tumors. For example, trisomy 12 is found to be common in both pluripotent p53 stabilization following DNA damage in bulge SC. The attenuated p53 stem cells and germ cells tumors, while aberrations in chromosome 18 arise activation is the consequence of a faster DNA repair activity mediated by a in both neural stem cells and neuroblastomas. Together, these data suggest

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higher non-homologous end joining (NHEJ) activity. Since NHEJ is an error protein-coding regions, large genomic regions harboring non-coding RNAs prone mechanism, this novel characteristic of adult SCs could be a double also need to be reprogrammed to a stem-cell like state. For example, recent edged sword for SCs as it mediates immediate survival at the expense of studies demonstrated that the Chr12qF1/Dlk1-Dio3 locus which harbors long-term maintenance of genomic integrity. We are currently investigating a large cluster of non-coding RNAs plays a critical role in determining the the long-term consequences of this characteristic in relation to ageing and differentiation potential of iPS cell lines. However, the mechanistic link cancer. between chromatin biology and ncRNA during cellular reprogramming has not been explored. During adult homeostasis bulge SCs are mostly quiescent whereas dur- ing organogenesis and tissue regeneration, hair follicle SCs divide more Our preliminary studies have identified a novel epigenetic mechanism for frequently. Since different DNA repair mechanisms are used at different controlling ncRNA expression, especially, at the Chr12qF1 locus. We further stages of the cell cycle, the relative importance NHEJ versus homologous demonstrate its function during cellular reprogramming. Using a ChIP-seq recombination (HR) may depend on the stages of SC ontogeny. To address approach, we discovered that a specialized histone variant protein, H2A.X this question, we performed conditional deletion of genes involved in (counts for ~1% of the total H2A in the differentiated cells), is targeted to HR at different stages of hair follicle development and/or activation. The critical regions for ES cell differentiation and reprogramming. Chr12qF1 is functional consequences of these experiments in term of SC maintenance, among the strongest loci where H2A.X imposes its effect since the non-cod- ageing and cancer will be presented. ing RNAs at this locus are mainly derived from TE. In parallel, the expression of ncRNA genes at Chr12qF1 is largely dependent on H2A.X, revealed by a This work was supported by the FNRS, the program d’excellence CIBLES of RNA-seq approach. Importantly, by a reconstitution approach, we showed the Wallonia Region, Fondation Contre le Cancer, the European Research that H2A.X plays a direct role in transcriptional regulation of the expres- Council (ERC) and the EMBO Young Investigator Program. sion of these ncRNAs. In-depth analysis further demonstrated that H2A.X plays a novel role in regulating high order chromatin structure at ncRNA locus. The underlying molecular mechanisms controlled by H2A.X will also Concurrent Session III Track D – be discussed. Finally, we extend our investigation to show that H2A.X plays an important role during iPS, especially at “resetting” the Chr12qF1 locus Epigenetic Programming of Stem Cells which is critical for the differentiation/developmental potential of the iPS cells. Saturday, June 18, 1:30 – 3:15 p.m. In summary, we found a novel function for histone variant in controlling ncRNA expression. These findings may have far-reaching impacts on cellular ROLE OF TET PROTEINS IN DNA reprogramming and differentiation. METHYLATION AND EMBRYONIC STEM CELL SELF-RENEWAL THE ROLE OF DNA METHYLATION IN Zhang, Yi REGULATING TRANSCRIPTOME IN MOUSE Howard Hughes Medical Institute and Dept of Biochemistry and Biophysics, University of North Carolina at Chapel Hill, Chapel Hill, NC, EMBRYONIC STEM CELLS USA Huang, Kevin1, Yu, Juehua1, Le, Thuc1, Pellegrini, Matteo2, Fan, Epigenetic modifications play important roles in diverse biological processes Guoping1 that range from regulation of gene expression, embryonic development, 1Dept of Human Genetics, UCLA, Los Angeles, CA, USA, 2Molecular, Cell stem cell reprogramming, and human diseases such as cancers. One of the and Developmental Biology, UCLA, Los Angeles, CA, USA epigenetic modifications is DNA methylation. Although enzymes respon- sible for DNA methylation have been well characterized, enzymes that re- DNA methylation is an epigenetic mechanism primarily associated with sponsible for active DNA demethylation in mammalian cells have remained gene silencing. However, recent genome-wide studies indicate DNA meth- a controversial topic. Recent studies have demonstrated that the Tet family ylation may possess additional properties. We generated mouse embryonic proteins can catalyze conversion of 5-methylcytosine to 5-hydroxymeth- stem (ES) cells that are deficient in all three major DNA methyltransfer- ylcytosine, a potential step toward DNA demethylation. Importantly, Tet1 ases via the Cre/loxP system. Mass spectrometry confirmed these triple plays an important role in embryonic stem cell self-renewal, maintenance, knockout (TKO) cells are virtually devoid of cytosine methylation. To survey and embryonic development. In my talk, I will discuss our effort in under- the RNA transcriptome in the absence of DNA methylation, we performed standing how Tet1 regulate these processes. RNA-Seq in control and TKO ES cells. Our data revealed dramatic changes in the RNA landscape in the absence of DNA methylation. For example, TKO cells show deregulation of genes involved in immune response and metabolism. Concurrently, we also observe genes with differential isoform HISTONE VARIANT PROTEIN H2A.X PLAYS A expression in TKO cells. By comparing the number of unique exon-exon NOVEL ROLE IN STEM CELLS junctions per gene between TKO and control ES cells, our data suggest

1 1 1 2 that DNA methylation does not have a single rule on modulating isoform Wu, Tao , Liu, Yifei , Drane, Pascal , Stadtfeld, Matthias , Kim, expression. Finally, we identify a subset of genes that show a switch in 1 2 1 Jeehee , Hochedlinger, Konrad , Xiao, Andrew transcript direction, suggesting DNA methylation plays roles in suppress- 1Yale Stem Cell Center and Department of Genetics, Yale School of ing strand-specific transcription. These genes are often low or moderately Medicine, New Haven, CT, USA, 2Massachusetts General Hospital Cancer expressed in both TKO and control cells. Taken together, our results demon- Center, Howard Hughes Medical Institute and Department of Stem Cell strated that DNA methylation plays multiple roles in regulating RNA tran- and Regenerative Biology, Harvard University, Boston, MA, USA scriptomes in mouse ES cells, ranging from gene silencing to RNA splicing. Recent advance in cellular reprogramming (induced pluripotent stem cells iPSc) presents a promising venue for regenerative medicine. Histone-medi- ated mechanisms play important roles in reversing the epigenetic land- scape during cellular reprogramming. Previous studies focused on histone modification pattern change at the protein-coding genes. In addition to

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CORD BLOOD-DERIVED NEURONS BY TET family enzymes convert 5-methylcytosine (5mC) to 5-hydroxymeth- ylcytosine (5hmC) in DNA. Here, we show that Tet1 and Tet2 are Oct4- ECTOPIC EXPRESSION OF SOX2 AND CMYC regulated enzymes that together sustain 5hmC in mouse embryonic stem Giorgetti, Alessandra1, Marchetto, Maria C.N2, Fazzina, cells (ESCs) and are induced concomitantly with 5hmC during reprogram- Raffaella1, Li, Mo2, Yu, Diana2, Mu, Yangling2, Gage, Fred H.2, ming of fibroblasts to induced pluripotent stem cells. ESCs depleted of Tet1 Izpisua Belmonte, Juan Carlos2 by RNAi show diminished expression of the Nodal antagonist Lefty1 and display hyperactive Nodal signaling and skewed differentiation into the 1 2 Stem Cell Bank, CMRB, Barcelona, Spain, Salk Institute for Biological endoderm-mesoderm lineage in embryoid bodies in vitro. In Fgf4- and Studies, La Jolla, San Diego, CA, USA heparin-supplemented culture conditions, Tet1-depleted ESCs activate The finding that the epigenome of differentiated cells can be reset to a the trophoblast stem cell lineage determinant Elf5 and can colonize the pluripotent state indicates that any somatic cell may potentially change or placenta in midgestation embryo chimeras. Consistent with these findings, reverse its already established developmental identity through the delivery Tet1-depleted ESCs form aggressive hemorrhagic teratomas with increased of appropriate instructive signals. Here we show the possibility of generat- endoderm, reduced neuroectoderm, and ectopic appearance of trophoblas- ing mature and functional neurons from Cord Blood (CB) cells without tic giant cells. Thus, 5hmC is an epigenetic modification associated with the reprogramming into a pluripotent state. In particular, we demonstrate that pluripotent state, and Tet1 functions to regulate the lineage differentiation the ectopic expression of two transcription factors (SOX2, c-MYC) as well potential of ESCs. as only one factor (SOX2) allows the generation of proliferating neural TET2 is a close relative of TET1, an enzyme that converts 5-methylcytosine progenitor cells starting from CB CD133+ cells. Given its important role (5mC) to 5-hydroxymethylcytosine (5hmC) in DNA. The gene encoding in neural stem cell activity, we first tested if only SOX2 was sufficient to TET2 resides at chromosome 4q24, in a region showing recurrent microde- induce the conversion of CB cells into neural progenitor cells. Three weeks letions and copy-neutral loss of heterozygosity (CN-LOH) in patients with after retrovirus infection, we observed the formation of a few colonies diverse myeloid malignancies. Somatic TET2 mutations are frequently ob- showing an iPS morphology, although they were negative for pluripotency served in myelodysplastic syndromes (MDS), myeloproliferative neoplasms markers (OCT4, NANOG, SSEA4). However, these colonies homogenously (MPN), MDS/MPN overlap syndromes including chronic myelomonocytic expressed multiple neural markers (TJU1, GFAP, MAP2, PAX6, NF) and we leukaemia (CMML), acute myeloid leukaemias (AML) and secondary called them CB-inducible neural progenitor cells (CB-iNPCs). Since, it is AML (sAML). We show here that TET2 mutations associated with myeloid known that c-MYC has a role in controlling self-renewal and proliferation of malignancies compromise catalytic activity. Bone marrow samples from neural progenitor cells, we therefore tested the neuron-inducing activity of patients with TET2 mutations displayed uniformly low levels of 5hmC in SOX2 in combination with c-MYC. The data showed that the presence of genomic DNA compared to bone marrow samples from healthy controls. c-MYC increased the efficiency 10 fold. In addition, we have demonstrated Moreover, small hairpin RNA (shRNA)-mediated depletion of Tet2 in mouse that CB-iNPCs were able to differentiate into functional mature neurons in haematopoietic precursors skewed their differentiation towards monocyte/ vitro and in vivo. The expression of the mature synaptic marker, synapsin, macrophage lineages in culture. There was no significant difference in and both excitatory (VGLUT-1) as well as inhibitory markers (GABA), indi- DNA methylation between bone marrow samples from patients with high cated that CB-derived neurons had the protein machinery necessary to fire 5hmC versus healthy controls, but samples from patients with low 5hmC action potentials. To investigate if CB-derived neurons had functional mem- showed hypomethylation relative to controls at the majority of differentially brane properties similar to neurons, we performed patch-clamp recording methylated CpG sites. Our results demonstrate that Tet2 is important for and calcium transient assays. The results confirmed that CB-derived neurons normal myelopoiesis, and suggest that disruption of TET2 enzymatic activity could form functional synapses and generate action potentials. Finally, an favours myeloid tumorigenesis. Measurement of 5hmC levels in myeloid in vivo assay, where CB-derived neurons were injected into dentate girus malignancies may prove valuable as a diagnostic and prognostic tool, to of mice hippocampus, demonstrated that these cells were able to engraft, tailor therapies and assess responses to anticancer drugs. differentiate as well as extend processes along the corpus callosum, one month after transplantation. These data suggest that the ectopic expression of SOX2 and c-MYC as well as only SOX2 can rapidly convert CB cells into function neurons. The possibility to generate functional neurons starting Concurrent Session III Track E – from CB cells, in an efficient and easy way, could offer a novel and powerful system for studying human cellular identity and plasticity. Cardiac & Muscle Stem Cells Saturday, June 18, 1:30 – 3:15 p.m. ROLE OF TET PROTEINS IN MOUSE RADIAL CONSTRUCTION OF AN ARTERIAL EMBRYONIC STEM CELLS AND MYELOID WALL TUMORIGENESIS Krasnow, Mark Koh, Kian Peng1, Ko, Myung Gon2, Huang, Yun2, Yabuuchi, Akiko3, Rao, Sridhar3, Sommer, Cesar A.4, Mostoslavsky, Gustavo4, Stanford University, Stanford, CA, USA Orkin, Stuart H.3, Jankowska, Anna M.5, Agarwal, Suneet3, Some of the most serious human diseases such as aortic aneurysm, coro- Maciejewski, Jaroslaw P.5, Daley, George Q.3, Rao, Anjana1 nary artery atherosclerosis and pulmonary hypertension are associated with altered size and structure of the arterial wall. Elucidating how arterial walls 1Immune Disease Institute and Program in Cellular and Molecular are built and maintained could aid understanding of these diseases, but little Medicine, Boston, MA, USA, 2La Jolla Institute for Allergy and is known about how the concentric layers of smooth muscle cells and outer Immunology, La Jolla, CA, USA, 3Division of Pediatric Hematology/ adventitial layer are assembled and patterned around endothelial tubes. Oncology, Children’s Hospital Boston and Dana-Farber Cancer Using histochemical, clonal, and genetic analysis in mice, here we show that Institute, Harvard Stem Cell Institute, Boston, MA, USA, 4Section of the pulmonary artery wall is constructed radially, from the inside out, by Gastroenterology, Department of Medicine, and Center for Regenerative two separate but coordinated processes. One is sequential induction and Medicine, Boston University School of Medicine, Boston, MA, USA, recruitment of successive cell layers from surrounding mesenchymal cells. 5Department of Translational Hematology and Oncology Research, Taussig The other is controlled invasion of outer layers by dividing inner layer cells, Cancer Institute, and Department of Hematologic Oncology and Blood through developmentally-regulated spindle reorientation and radial migra- Disorders, Cleveland Clinic, Cleveland, OH, USA 90 www.isscr.org Final Program

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tion. We propose that an endothelial-derived radial patterning factor con- phenotype is not evident in mdx mice because the long telomeres result in a trols and coordinates the two processes, and our genetic data demonstrate stem cell reservoir that suffices to repair the chronically damaged dystro- that PDGF-B, previously implicated in arterial wall development, is not it. phin-deficient myofibers throughout the lifetime of the mouse. To test this Identification of the factor would reveal the molecular basis of arterial wall hypothesis we mated dystrophin deficient mice (mdx) with telomerase patterning and suggest new therapeutic targets for arterial diseases. deficient mice (mTRKO). The double mutant mice (mdx/mTRKO) exhibit all of the hallmarks of the of DMD: markedly elevated creatine kinase levels, severe loss of muscle force, inability to run on a treadmill, kyphosis, muscle IN VIVO REPROGRAMMING OF membrane disruption, progressive wasting of limb and diaphragm muscles, culminating in early death. As shown previously in humans, the muscle MURINE CARDIAC FIBROBLASTS INTO stem cells in this mouse model exhibit reduced proliferative potential, both CARDIOMYOCYTES in vitro and in vivo, and impaired ability to engraft in and regenerate mdx/ mTRKO muscles upon transplantation. In contrast, transplantation of WT Qian, Li, Huang, Yu, Foley, Amy, Fu, Jidong, Srivastava, Deepak muscle stem cells (MuSC) into mdx/mTRKO mice ameliorates the dystro- Gladstone Institutes, San Francisco, CA, USA phic phenotype histologically. Together, these results indicate that the com- Heart failure affects millions worldwide and is a progressive disease. The bination of the ongoing damage due the structural defect of dystrophin- human heart has limited endogenous regenerative capacity and is thus deficiency that leads to muscle degeneration together with the progressive a target for novel regenerative medicine approaches. Cardiac fibroblasts exhaustion of functional MuSC to repair the chronically induced damage comprise approximately 50% of cells in the mammalian heart and contrib- leads to the manifestation of the dystrophic phenotype. These findings ute to scar formation upon cardiac damage. Our recent report showing support the hypothesis that, in humans, DMD is initiated by the structural direct reprogramming of fibroblasts into cardiomyocyte-like cells by defined genetic defect but develops progressively into an exhaustion of stem cells factors in vitro raises the possibility that endogenous cardiac fibroblasts with regenerative potential due to telomere deficiency. could serve as a potential source of new cardiomyocytes for regenerative therapy. Here, we show that resident fibroblasts can be reprogrammed into cardiomyocyte-like cells in the murine heart by local delivery of Gata4, ENGINEERED SKELETAL MUSCLE CONTAINS Mef2c and Tbx5 (GMT). We utilized a transgenic mouse with Cre-re- FUNCTIONAL SATELLITE CELL NICHES combinase under control of the fibroblast-enriched promoter of Periostin, normally excluded from cardiomyocytes, crossed with the R26R-lacZ mouse CAPABLE OF MUSCLE REGENERATION IN to lineage trace the emergence of newly reprogrammed cardiomyocytes. VITRO Retroviral introduction of (GMT) into the infarct/border zone after coronary ligation resulted in the generation of numerous lacZ-positive cells that also Tiburcy, Malte, Markov, Aleksej, Zimmermann, Wolfram H. expressed cardiac-specific sarcomeric genes and gap-junction proteins, Pharmacology, Georg-August-University Goettingen, Goettingen, Germany suggesting electrical coupling of new myocytes with neighboring myocytes. Introduction: Satellite cells reside in defined niches in skeletal muscle. They Furthermore, in vivo reprogramming of cardiac fibroblast into cardiomyo- have high regenerative capacity which may be reduced with age and in my- cytes in a mouse myocardial infarction (MI) model attenuated infarct size opathies. Analysis of satellite cell function in vitro is limited as they quickly and rescued cardiac dysfunction, even 12 weeks after injury. These results differentiate and lose their phenotype. We hypothesized that (1) skeletal indicate that a significant fraction of endogenous cardiac fibroblasts can be muscle (ESM) with functional satellite cell niches can be engineered in vitro diverted to generate new cardiomyocytes and that introduction of cardiac and (2) exploited to study skeletal muscle regeneration. reprogramming factors into cardiac fibroblasts in vivo can improve cardiac function after myocardial infarction. Methods and Results: We generated engineered skeletal muscle (ESM) from rat myoblasts, matrigel, and collagen (1.25x106 cells/ESM). ESM displayed highly differentiated muscle syncytia with typical skeletal muscle proper- ADULT STEM CELL EXHAUSTION DUE TO ties (tetanic contractions, functional acetylcholine receptors). In contrast to conventional 2D culture, ESMs exhibited Pax7-positive cells in satellite cell- SHORT TELOMERES IN CHRONICALLY characteristic niches: m-cadherin+, caveolin+ cells adjacent to muscle fiber DAMAGED DYSTROPHIN-DEFICIENT MDX/MTR and underneath laminin+ basal membrane. Immune staining indicated that the majority of Pax7 positive satellite cells was quiescent (Ki67-, MyoD- MICE MIMICS HUMAN DUCHENNE MUSCULAR ). We next investigated whether satellite cells would entail regenerative DYSTROPHY capacity using three independent assays: (i) cardiotoxin injury - treatment

1 2 1 with 25 µg/ml cardiotoxin caused muscle cell destruction and contractile Mourkioti, Foteini , Sacco, Alessandra , Blau, Helen M. failure after 48 hrs. However, Pax-7 positive satellite cells were spared. 1Microbiology and Immunology, Stanford University, Stanford, CA, USA, After 7 days of recovery we found a partial regeneration of contractile force 2Muscle Development and Regeneration Program, Sanford Children’s (33±4% of control, n=6-11/group). This was paralleled by a regenera- Health Research Center, Sanford-Burnham Medical Research Institute, CA, tive gene expression pattern (increased Pax7, MyoD, embryonic myosin USA heavy chain [eMHC] transcripts, n=4; p<0.05). Immune staining revealed Duchenne muscular dystrophy is a devastating genetic disease caused by proliferation and differentiation of activated Pax7-positive satellite cells with a mutation in the dystrophin gene which occurs in ~1/3500 males. The regenerating muscle fibres (eMHC-positive); (ii) mechanical injury - defined absence of a functional dystrophin leads to progressive degeneration of regions of circular ESMs were injured by repeated short squeezes with for- skeletal muscles, loss of regenerative potential, and death of patients at a ceps while others were spared (n=6). Gene expression showed a differential young age. A major limitation in the study of the pathogenesis of DMD and response with upregulation of Pax-7, MyoD, and eMHC only in the injured ability to test therapeutic strategies is the absence of an adequate mouse arm (p<0.05); (iii) derivation of functional satellite cells from ESMs - we model of the disease that exhibits the severe clinical symptoms seen in isolated satellite cells from ESM and after satellite cell expansion (5 days) patients. Indeed, the skeletal muscles of the mouse model with the same generated new ESMs. One ESM generated enough cells to construct 6 new genetic deficiency (mdx mice), are capable of repeated cycles of degenera- force-generating ESMs. tion and regeneration and regenerative capacity does not appear to be Conclusion: We demonstrate a novel technique to engineer skeletal muscle exhausted. A major difference between mice and humans is the length of (ESM) with functional satellite cell niches in vitro. We further provide evi- their telomeres, which is far greater in mice. We postulated that the DMD 91 ISSCR 9th Annual Meeting www.isscr.org

Detailed Program and Abstracts – Saturday, June 18

dence for regenerative activity of engineered satellite cells niches in models A GENOME-WIDE SIRNA SCREEN IDENTIFIES of muscle injury and after satellite cell re-isolation, providing the ground for a further exploitation of ESMs in studies of mechanisms underlying skeletal SELECTIVE INHIBITORS OF BASAL-LIKE muscle injury and regeneration as well as potential pharmacological or BREAST TUMOR-INITIATING CELLS molecular treatment of myopathies. Lieberman, Judi1, Petrocca, Fabio1, Altschuler, Gabriel2, Tan, Mynn1, Johnston, Sean3, Rudnicki, Stewart3, Richardson, Andrea4, 3 2 5 PROGENITOR CELLS IN ZEBRAFISH Shamu, Caroline , Hide, Winston , Ince, Tan 1Immune Disease Institute, Harvard Medical School, Boston, MA, USA, ORGANOGENESIS 2Harvard School of Public Health, Boston, MA, USA, 3ICCB-Longwood, Stainier, Didier Harvard Medical School, Boston, MA, USA, 4Brigham and Women’s Hospital, Boston, MA, USA, 5University of Miami Miller School of University of California, San Francisco, San Francisco, CA, USA Medicine, Miami, FL, USA ABSTRACT NOT AVAILABLE AT TIME OF PRINTING Basal-like breast tumors are an especially aggressive group of breast cancer, frequently occurring in patients carrying BRCA1 genetic mutations. These tumors rapidly acquire resistance to chemotherapy and are refractory to Plenary VIII – Regulatory Networks of Stem Cells endocrine therapy and HER-2 inhibitors. Breast primary epithelial cells give rise to different cancer cell types after transformation with the same genetic Saturday, June 18, 4:25 – 6:40 p.m. elements depending on growth medium (Ince and Weinberg). BPLER cells have the phenotype of bipotent early epithelial progenitor cells, while HM- TRANSCRIPTIONAL CONTROL OF EMBRYONIC LER cells are more differentiated myoepithelial cells. Orthotopic injection of 50 BPLER cells in immunodeficient mice uniformly gives rise to tumors that STEM CELLS closely resemble human basal-like breast tumors, while 5x104 HMLER cells Young, Richard A. do not initiate tumors. Therefore BPLER cells are highly enriched for breast tumor-initiating cells (BT-IC). We performed a genome-wide siRNA screen Whitehead Institute for Biomed Research and MIT, Cambridge, MA, USA to identify factors on which tumor-initiating BPLER cells selectively depend Discovering how transcriptional regulation establishes and maintains gene for survival. We identified a set of 154 genes whose silencing selectively expression programs in mammalian cells is important for understanding affects survival and proliferation of these BT-ICs. This set of genes is highly the control of cell state, the process of development and the mechanisms expressed in breast cancer relative to normal breast tissue and expression involved in various diseases. I will discuss key features of transcriptional correlates with poor prognosis in breast cancer. Of these 154 genes, 15 regulatory circuitry in embryonic stem cells (ESCs), including the roles that genes regulate various aspects of the ubiquitin/proteasome system, which is specific transcription factors and chromatin regulators play in the establish- activated in human primary basal-like breast tumors. Silencing these genes ment and maintenance of pluripotency and self-renewal. We have identi- triggers apoptosis in basal-like BPLER cells, but not HMLER cells or normal fied novel mechanisms that link gene expression and chromatin structure, breast epithelial progenitors. BT-ICs are also selectively sensitive to low-dose and other mechanisms that help remodel transcriptional regulatory circuitry proteasome inhibitor drugs. Since proteasome inhibitors are already in the during differentiation. Advances in our understanding of the regulation of clinic, these findings may be rapidly investigated for breast cancer therapy gene expression programs are providing new insights into the mechanisms in a clinical setting. that control cell state and development and, when altered, contribute to a variety of diseases.

REGULATORY NETWORKS IN STEM CELLS AND CANCER Orkin, Stuart H. Dana-Farber Cancer Institute, Children’s Hospital Boston, Howard Hughes Medical Institute, Harvard Stem Cell Institute, Harvard Medical School, Boston, MA, USA Polycomb group proteins constitute epigenetic repressors of cell fates and differentiation. We have focused on the composition and function of Poly- comb Repressive Complex-2 (PRC2) in embryonic stem (ES) cells, the he- matopoietic system, and oncogenesis. In this presentation I will first review findings regarding the composition of PRC2 in ES cells, the identification of Jarid 2(Jmj) as a critical component, and the PRC2 transcriptional network. In our studies we have used knockout and conditional gene targeting to ex- amine the roles of PRC2 by formal genetics. Recent findings regarding the requirements for PRC2 components in hematopoietic stem cells (HSCs) and hematopoietic differentiation will be presented. The overexpression of PRC2 in various cancers has suggested that PRC2 is oncogenic, and might repre- sent a therapeutic target. I will present some of our efforts to determine the contribution of PRC2 to both hematopoietic and solid tumor malignancies. Overall, our findings provide insights into the complex roles of Polycomb proteins in stem cell biology and cancer.

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Detailed Program and Abstracts – Saturday, June 18

The Anne McLaren Memorial Lecture THE NEURAL CREST, A PLURIPOTENT STRUCTURE OF THE VERTEBRATE EMBRYO Le Douarin, Nicole M. Academie Des Sciences, Paris, France I started to work on the neural crest (NC) in 1969, after I devised a tech- nique that allowed following the migration of cells in the avian embryo in ovo. In this technique, chimeras were constructed between two species of birds , the chick and the japanese quail, whose cells could be easily recog- nized by the structure of their nucleus, thus providing a stable, non-diffus- ible, cell marker. With my co-workers, we constructed a fate map of the NC derivatives along the neural axis at a stage preceding the onset of NC cell (NCC) emigration and we described the pathways followed by the NCC to reach their destination. The NCC are a very invasive cell type and the NC is a highly pluripotent structure which gives rise to the peripheral nervous sys- tem, melanocytes, endocrine cells and, at the cephalic level, the entire facial skeleton, part of the cranial bones and of most of the connective tissues of the head. At the trunk level, in contrast, no cells of the mesenchymal and skeletal types are provided by the NC in vivo in higher vertebratees. The second phase of this work entailed several analyses: 1- Investigation of the developmental potentialities of individual cephalic NCC from both the cephalic and trunk levels. Clonal cultures of NCC, at the time they start migrating, showed that early NCC are endowed with broad developmental capacities: a large proportion of them gave rise to clones containing both neurons, glia, melanocytes together with chondroblasts, osteoblasts and myofibroblasts, thus showing that the segregation between mesodermal and ectodermal derivatives did not take place in the NC. More- over these experiments showed that pluripotent stem cells exist in the NC. 2- Since the truncal NC (TNC) of lower vertebrates generates mesenchymal tissues, such as the dermal skeleton of fish caudal fin, the question arose as to whether the ability of NC cells to yield mesenchyme has been lost in the trunk during evolution and whether it can still be elicited in amniotes under specific conditions. We succeeded in devising culture conditions in which avian TNC yielded “mesenchymal” cell types such as osteoblasts and myofibroblasts. 3- In our search for cytokines which trigger the proliferation and/or the differentiation of the various types of NC derivatives, we found that Endo- thelin-3 (Et-3) strongly favours the development of melanocytes in primary NCC cultures. Moreover, treatment with ET-3 was able to reprogram dif- ferentiated melanocytes into a type of pluripotent NC stem cell yielding, in clonal cultures, myofibroblasts, adipocytes, glial cells, melanocytes and stem cells that could be propagated in culture. In this example, the reprogram- mation of differentiated cells into pluripotent stem cells could be obtained by the induction of cell proliferation.

93 ISSCR 9th Annual Meeting www.isscr.org

Innovation Showcases

Thursday, June 16, 11:45 a.m. – 12:15 p.m. Thursday, June 16, 11:45 a.m. – 12:15 p.m. Integrated Solutions for the Expansion and Multiparametric Analysis with Mass Cytometry Analysis of Human Mesenchymal Stem Cells Dmitry Bandura and Olga Ornatsky Abel Hastings DVS Sciences Inc. BD Biosciences Room 716 A/B Room 801 A/B Mass Cytometry is a revolutionary technology based on atomic mass Fulfilling the clinical promise of mesenchymal stem cells (MSCs) in cell spectrometry that addresses flow cytometry applications, but extends the therapy will require solutions to reliably expand these cells. BD Mosaic™ capability to many simultaneous parameters. The CyTOF™ Mass Cytom- hMSC SF, a novel chemically-defined serum-free MSC expansion medium, eter “reads” stable isotope tags attached to antibodies using the currently and BD Falcon™ Cell Culture MultiFlask, a new multi-layer culture flask, available 32 different MAXPAR™ labeling kits. Because there are up to 100 were used for the rapid and efficient expansion of MSCs. Cells cultured in stable isotopes, and the mass spectrometer provides exquisite resolution this system maintained their immunophenotype, as demonstrated with the between detection channels, many parameters can be measured as easily BD Stemflow™ MSC Analysis Kit, as well as their multipotency and immu- as one. nosuppressive capabilities. Thursday, June 16, 11:45 a.m. – 12:15 p.m. Thursday, June 16, 11:45 a.m. – 12:15 p.m. Innovation in Scotland: Novel Stem Cell Tools STEMdiff™ APEL™, A Defined Animal Product and Technologies Free Medium for the Growth Factor Directed David C. Hay, Verna McErlane and Jo Mountford Differentiation of Pluripotent Stem Cells Scottish Development International Elizabeth Ng Room 718 A/B STEMCELL Technologies Inc. Scotland has long been associated with scientific innovation and is a world Room 701 A/B leader in stem cell R&D. Overview of Scotland’s stem cell landscape fol- lowed by Fibromed, developing high fidelity liver models using pluripotent The efficient and reproducible differentiation of pluripotent human embry- stem cells; Sistemic’s recently launched SistemQC ™ service, to charac- onic and induced pluripotent stem cells remains one of the great challenges terise and monitor the QC of stem cell lines for identity, purity, safety and that must be met before the full potential of these cells can be realized. We differentiation staging using microRNA-based dynamic fingerprinting; and will present data showcasing the differentiation of human pluripotent cells Glasgow University’s innovative research to produce synthetic whole blood to multiple cell lineages in a growth factor dependent fashion in STEMdiff™ from ESC. APEL™ (Ng et al., 2008), a defined animal product free differentiation medium that contains recombinant human proteins. Thursday, June 16, 12:30 – 1 p.m. Thursday, June 16, 11:45 a.m. – 12:15 p.m. Induced Pluripotent Stem Cells for Research, Use of a Novel Synthetic Surface for the Drug Screening and Toxicity Testing Derivation, Proliferation and Differentiation of Jayne Hesley iPS Cells and Their Progeny Molecular Devices Inc. Michael J. Young Room 801 A/B Corning, Inc. Human iPS-derived cells are an ideal model system for testing both protective and toxic effects of drug candidates. We demonstrate how the Room 714 A/B combined use of high-content imaging, automated patch clamp, and kinetic We describe the use of the Corning® Synthemax™ surface to support fluorescence techniques provide a broad assay set to identify candidates mouse and human iPS cells during reprogramming and proliferation stages, early in drug discovery. as wells as through the process of differentiation into retinal neurons. Com- pared to extracellular matrix-coated surfaces, plating on Synthemax surface Thursday, June 16, 12:30 – 1 p.m. resulted in enhanced cellular adhesion. Synthemax surface provides an ideal surface for the xeno-free production, culture, and differentiation of adult Large Particle Flow Cytometry for Cells and mouse and human fibroblast derived iPSCs. These findings demonstrate the Cell Clusters in Stem Cell Research potential utility of this surface for the production of clinical grade retinal neurons for transplantation and induction of retinal regeneration. Rock Pulak Union Biometrica Inc. Room 701 A/B Cells growing in clusters communicate with each other and behave differ- ently than cells grown as monolayers or in suspension. These interactions are likely to be important for proper function. Union Biometrica Large Particle Flow Cytometers automate the analysis, sorting and dispensing of objects too big (10-1500μm) or too fragile for traditional cytometers including those studied by stem cell researchers such as embryoid bodies, neurospheres and other spheroids. 94 www.isscr.org Final Program

Innovation Showcases

Thursday, June 16, 12:30 – 1 p.m. Friday, June 17, 11:45 a.m. – 12:15 p.m. Isolation of Single Cells from SPKLS StemAdhere™: A Defined and Entirely Human Sub-populations by Flow Cytometry for Substrate for the Culture of hESCs and hiPSCs Downstream Single Cell Gene Expression Stephen A. Duncan Profiling STEMCELL Technologies Inc. Gil Reinin Room 801 A/B BD Biosciences StemAdhere™ is developed by Primorigen Biosciences and is a defined and Room 714 A/B entirely human recombinant protein substrate that supports the mainte- nance and expansion of human embryonic stem cells (hESCs) and human The combination of flow cytometry and single cell gene expression forms induced pluripotent stem cells (hiPSCs) cultured in defined, feeder-indepen- a powerful tool for stem cell research. HSC subtypes were identified on the dent maintenance medium. StemAdhere™ can reproducibly support the basis of Hoechst dye efflux and SPKLS surface staining using the BD Stem- long term passaging of hESCs and hiPSCs while maintaining high expres- flow™ Mouse Hematopoietic Stem and Progenitor Cell Isolation Kit. Single sion of pluripotency markers. This commercial tutorial will discuss the key cells from SP sub-populations, My-HSCs and Ly-HSCs, were isolated using a performance criteria that make StemAdhere™ a desirable alternative to BD FACSAria™III. Differences in gene expression were found between the traditional matrices. two populations using the Fluidigm BioMark™ HD System. Friday, June 17, 11:45 a.m. – 12:15 p.m. Thursday, June 16, 12:30 – 1 p.m. Multiplex Gene-Expression Assay for Human Reproducible Derivation of Integration-Free Induced Pluripotent Stem Cells (iPSCs) Using Human iPS Cells Using mRNA GexP Genetic Analysis System Brad Hamilton Bee-Na Lee Stemgent Beckman Coulter Inc. Room 716 A/B Room 701 A/B Until recently, the generation of human iPS cells without genomic modifi- cation had proven to be a highly inefficient and difficult process. In 2010, Monitoring gene expression level of the reprogramming genes and stem- Warren et al demonstrated that the use of mRNA for cellular reprogram- ness biomarker genes in induced pluripotent stem cells (iPSCs) is an efficient ming was not only feasible, but also a highly productive methodology for method to identify clones that are potentially undergoing certain differen- generating integration-free human iPS cells. This presentation will highlight tiation pathways. We designed a multiplex gene assay containing endoge- the internal validation of a new line of Stemgent mRNA reprogramming nous SOX2, OCT4, KLF4, cMYC, LIN28, NANOG genes and its counterpart products for the reliable and consistent generation of integration-free trans-genes to provide a cost-effective molecular testing tool in determina- human iPS cells. tion of desirable clones during the early stage of stem cell research/study.

Thursday, June 16, 12:30 – 1 p.m. Friday, June 17, 11:45 a.m. – 12:15 p.m. iPS Cell Repository for Human Tissue and New Techniques to Control Pluripotent Stem Disease Models Cell Differentiation Will Rust Andreas Bosio ATCC Miltenyi Biotec GmbH Room 718 A/B Room 714 A/B An iPS cell repository has been created at the ATCC to accession and Various protocols for differentiation of stem cells into specific neural globally distribute highly qualified, standardized cell lines reprogrammed subpopulations have been reported. We have optimized these protocols via expression of OSKM. This tutorial showcases the growing collection of by implementing magnetic cell sorting steps. The enrichment of mouse ES cells derived from normal and diseased tissues without gene integration. cell-derived neuronal precursor cells streamlines the in vivo differentiation A novel media formulation is introduced which simplifies pluripotent cell towards neurons and increases the number of target cells after grafting. research. Also outlined is how the ATCC can effectively enable researchers Likewise, magnetic enrichment of pluripotent human iPS cells prior to requiring models of human tissue and disease. neural induction circumvents the need for pre-expansion in conditioned medium, saving time, and more importantly, leading to predictable differen- tiation results.

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Innovation Showcases

Friday, June 17, 11:45 a.m. – 12:15 p.m. Friday, June 17, 12:30 – 1 p.m. Isolation of Mouse Hematopoietic Stem Cell A Leading Global Stem Cell Research and Side Populations Using SPKLS and Post-Sort Preclinical R&D Partner Confirmation Using Single-Cell Gene Expression John Listello Ken Livak and Alain Mir Sigma Life Science Fluidigm Room 701 A/B Room 716 A/B The development, standardization and application of stem cells for regen- Fluidigm will demonstrate a powerful method that combines single-cell erative medicine presents some of the most exciting opportunities to better FACS sorting with Fluidigm high-throughput real-time PCR technology. understand and treat diseases. Understanding the biology of pluripotency, This approach permits correlation of flow cytometry data with expression cellular expansion, and differentiation requires a great deal of research and of hundreds of genes in single cells. Single cells were isolated and individu- partnerships within the global research community. Sigma is taking an ally screened for mRNA expression of hundreds of genes. This technique active role in the R&D of key tools and content with a number of academic permits correlation of protein expression data with comprehensive gene and commercial partners and strives to accelerate progress in this area. expression profiles of single cells. Friday, June 17, 12:30 – 1 p.m. Friday, June 17, 11:45 a.m. – 12:15 p.m. Integrated Cell-Processing Device for Multiparameter Flow Cytometry and Automated Manufacturing of GMP-Compliant Bioimaging: Tools for Identifying Unique Stem Cell Products Immunophenotypes to Isolate and Analyze Stefan Miltenyi Sub-populations of Stem Cells and their Miltenyi Biotec GmbH Derivatives Room 714 A/B Christian Carson Cellular therapies are being investigated for an ever-increasing range of BD Biosciences applications. We have developed a new integrated cell-processing device in order to automate and standardize the complete manufacturing process Room 718 A/B of cellular products, including GMP-compliant cell separations as well as BD LyoplateTM Screening Panels are the first 96-well plated based, antibody pre- and post-separation handling steps, such as fractionation of starting screening panels available for the high-content screening of cell populations material, cell wash, target cell labeling, cell culture, volume reduction, and by imaging or flow cytometry. This tutorial will illustrate how the discovery final product formulation. As a first protocol, fully automated clinical-scale of unique immunophenotypic signatures enables the identification and separation of CD133+ cells from bone marrow aspirates has been estab- isolation of distinct cell populations from heterogeneous cell cultures. We lished. will demonstrate how this screening approach was used to isolate near pure populations of NSC, glia and neurons derived from pluripotent stem cells. Friday, June 17, 12:30 – 1 p.m.

Friday, June 17, 12:30 – 1 p.m. A Highly Efficient RNA Transfection Reagent for the Manipulation of Cell Fate Ex Vivo Expansion of Stem/Progenitor Cells Using Defined, Serum-Free Systems Kerry P. Mahon Stemgent Jessie H.-T. Ni Room 716 A/B R&D Systems Inc. The ability to influence cellular development and protein expression through Room 801 A/B the introduction of foreign genetic material is an effective and widely used Optimal stem/progenitor cell proliferation in vitro critically depends on research tool. Currently, manipulation of cell fate via RNA based meth- qualities of growth factors, basal media and, when applied, culture matrix ods is limited by the lack of suitable methods to deliver such material to used. A range of media, supplements and culture matrix have been devel- primary stemfect cell types. Here, we demonstrate the efficacy of the RNA oped by the stem cell group in R&D Systems and will be introduced in this Transfection Kit for a variety of RNA subtypes into a range of cells including showcase that would allow researchers to achieve ex vivo expansions of fibroblasts, hES and hiPS cells. pluripotent, mesenchymal and neural stem cells under defined, serum-free conditions.

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Innovation Showcases

Friday, June 17, 12:30 – 1 p.m. Saturday, June 18, 11:45 a.m. – 12:15 p.m. Multiparameter Flow Cytometry: Tools for NutriStem™ hESC/iPSC XF- The First Universal Isolating Pluripotent Stem Cells and Analyzing Low Protein Xeno Free Media to Culture Endoderm, Ectoderm and Mesoderm Lineages Pluripotent Stem Cells Nil Emre Michal Amit, PhD BD Biosciences BI - Biological Industries Room 718 A/B Room 716 A/B BD Biosciences has developed validated multicolor flow cytometry assays With the goal of “humanization” and “standardization” of the culturing for a wide range of stem cell types: This tutorial will demonstrate how BD environment for pluripotent stem cells, NutriStem™ was co-developed by StemflowTM Kits enable the: scientists from Technion and Biological Industries. NutriStem™ has been • consistent isolation of highly-pure, viable populations of hESCs validated in peer scientific publications, and meets two challenges: creation • analysis of cell surface and intracellular marker expression in pluripo- of a cell culture environment that approximates the physiological xeno-free tent stem cells and ectoderm, endoderm and mesoderm lineages (XF) environment, and development of a media that maintains pluripotency • analysis of multiple differentiation time-points in a single sample or without background differentiation in both feeder- dependent (FD) and multiple samples simultaneously when multiplexed with Fluorescent feeder- free (FF) cultures. Cell Barcoding methods Saturday, June 18, 12:30 – 1 p.m. Saturday, June 18, 11:45 a.m. – 12:15 p.m. Micro RNA and Epigenetic Regulation of Neural Advances in Reprogramming Efficiency and Stem Cell Differentiation Culture of iPS Cells Ron Hart, PhD Vi Chu, PhD Life Technologies EMD Millipore Room 714 A/B Room 701 A/B The fate of neural stem cells can be influenced by growth factor gradients, EMD Millipore’s STEMCCA lentivirus reprogramming kits enable the easy, cell-cell contact during migration, and endogenous programming, including efficient and convenient generation of human and mouse induced pluripo- transcription factor networks, chromatin modifications and microRNAs. A tent stem (iPS) cells. The STEMCCA kits use a single polycistronic lentiviral genome-wide approach utilizing ChIP-seq and microRNA-seq shows that vector to improve efficiency and reduce the number of viral integrations. differentiation requires both dynamic response to external stimuli and an We will discuss advances in both human and mouse reprogramming, endogenous mechanism for stabilization to be able to transition from one Cre-mediated excision of the exogenous reprogramming transgenes, and cell type to another. Our goal is to identify modes of precursor program- optimal cell culture and characterization of iPS cells. ming and how they can be altered.

Saturday, June 18, 11:45 a.m. – 12:15 p.m. Xeno-Free Derivation and Maintenance of Pluripotent Cell Lines Jordan E. Pomeroy Life Technologies Room 714 A/B Future clinical application of pluripotent cells is dependent upon standard- ized, defined growth conditions that adhere to regulatory guidelines. Eliminating xenobiotics from culture conditions is one step towards produc- tion of cell lines for clinical use. We have established a “xeno-free” culture system that combines human extracellular matrix proteins, human feeders, and a Knockout SR XenoFree CTS based growth medium (Invitrogen) for the derivation and maintenance of pluripotent cell lines.

97 ISSCR 9th Annual Meeting www.isscr.org

Poster Floor Plan

Totipotency/Early Embryo Cells Mesenchymal Mesenchymal Pre-Clinical and Clinical Germline Technologies for Tissue Hematopoietic Cell Lineage Stem Cell Applications of Neural Cell Cancer Cells Cells Stem Cell Research Engineering Stem Cells Analysis Di erentiation Mesenchymal Cells 2350 2250 2251 2270 2271 2290 2291 2310 2311 2011 2030 2031 2050 2051 2070 2071 2090 2091 2110 2111 2130 2131 2150 2151 2170 2171 2190 2191 2210 2211 2230 2010 2450 2451 2470 2471 2490 2491 2510 2511 2530 2531 2550 2551 2570 2571 2590 2591 2351 2370 2371 2390 2391 2410 2411 2430 2431 2330 2331 2231

745 844 845 944

743 842 843 942

741 840 841 940 2241 2260 2261 2280 2281 2300 2301 2320 2240 2020 2021 2040 2041 2060 2061 2080 2081 2100 2101 2120 2121 2140 2141 2160 2161 2180 2181 2200 2201 2220 2221 2001 2321 2340 2341 2440 2441 2460 2461 2480 2481 2500 2501 2520 2521 2540 2541 2560 2561 2580 2581 2600 739 838 839 938 2360 2361 2380 2381 2400 2401 2420 2421

1291 1300 3001 3020 1290 1281 Other 3040 3021 1271 1280 433 532 533 632 832 833 932 Neural Cell 20' Chromatin in 3041 3060 1270 1261 Stem Cells 3080 3061 1251 1260 331 430 431 530 531 630 63 733 831 930 931 1030 20' 3081 3100 1250 1241 3120 3101 329 428 629 728 729 830 829 928 929 1028 1231 1240 Muscle Cells 20' 20' 20' 20' 3121 3140 1230 1221 iPS Cells 3160 3141 427 526 527 626 1211 1220 Ethics 325 724 824 825 924 925 1024 3161 3180 20' 20' 20' 1210 1201 50' 3200 3181 Regeneration Epithelial Cells 1191 1200 323 422 623 722 723 823 922 923 1022 3201 3220 1190 1181 Eye or Retinal Cells 20' (not skin) Mechanisms 3240 3221 1171 1180 820 3241 20' 20' 20' 720 721 821 920 20' 1020 3260 1170 1161 Intestinal/Gut Cells Reprogramming 3280 3261 1151 1160 319 418 619 718 719 818 819 918 919 1018 3300 Intern 3281 1150 1141 Epidermal Cells 60' Embryonic Stem Cells 3320 3301 Endothelial Cells/ Clinical Application 1131 1140 715 814 815 914 3321 3340 1130 1121 Hemangioblasts 3360 3341 1111 1120 313 413 712 713 812 813 912 1012 3361 3380 1110 1101 20' 30' 30' 3400 3381 311 410 710 711 811 910 1091 1100 20' 20' 3401 3420 1090 1081 Cardiac Cells 20' 3440 3421 309 408 611 708 709 810 809 908 909 1008 1071 1080 20' 20' 20' Embryonic Stem Cell 3441 3460 1070 1061 Di erentiation 3480 3461 407 506 507 EXH Lung Cells SER 1051 1060 305 404 504 605 704 705 804 805 1004 3481 3500 1050 1041 20' 20' 20' 3520 3501 1031 1040 30Liver3 Cells 403 502 503 603 702 703 802 803 Embr902 yonic90 3Stem1002 Cell 20' 20' 3521 3540 1030 1021 Pluripotency 3560 3541 1011 1020 30Pancreatic1 400 Cells401 500 501 600 601 700 701 800 801 900 901 1000 3561 3580 1010 1001 3600 3581 Main Entrance Poster Session Schedule Wednesday, June 15 Thursday, June 16 Friday, June 17 Saturday, June 18 3:30 – 4:00 p.m. 11:00 a.m. – 8:00 p.m. 11:00 a.m. – 8:00 p.m. 11:00 a.m. – 4:00 p.m. All Posters Put On Display Posters Open for Viewing Posters Open for Viewing Posters Open for Viewing

4:00 – 8:00 p.m. 6:00 – 8:00 p.m. 6:00 – 8:00 p.m. 4:00 p.m. MAIN Posters Open for Viewing Poster Presentation Reception Poster Presentation Reception Posters Dismantle ALL ODD NUMBERED ALL EVEN NUMBERED ENTRANCE POSTERS PRESENTED POSTERS PRESENTED 98 www.isscr.org Final Program

Poster Floor Plan

Totipotency/Early Poster Boards by Topic Embryo Cells Mesenchymal Mesenchymal Pre-Clinical and Clinical Posters 1001 – 1300 Germline Technologies for Tissue Hematopoietic Cell Lineage Stem Cell Applications of Pancreatic Cells 1001 – 1020 Cancer Cells Cells Stem Cells Analysis Di erentiation Neural Cell Stem Cell Research Engineering Mesenchymal Cells Liver Cells 1021 – 1048 Lung Cells 1049 – 1060 2350 2250 2251 2270 2271 2290 2291 2310 2311 2011 2030 2031 2050 2051 2070 2071 2090 2091 2110 2111 2130 2131 2150 2151 2170 2171 2190 2191 2210 2211 2230 2010 2450 2451 2470 2471 2490 2491 2510 2511 2530 2531 2550 2551 2570 2571 2590 2591 2351 2370 2371 2390 2391 2410 2411 2430 2431 2330 2331 2231 Cardiac Cells 1061 – 1106 Endothelial Cells/ Hemangioblasts 1107 – 1140

745 844 845 944 Epidermal Cells 1141 – 1155

743 842 843 942 Intestinal/Gut Cells 1156 – 1165 Epithelial Cells (not skin) 1166 – 1177 741 840 841 940 Eye or Retinal Cells 1178 – 1204 2241 2260 2261 2280 2281 2300 2301 2320 2240 2020 2021 2040 2041 2060 2061 2080 2081 2100 2101 2120 2121 2140 2141 2160 2161 2180 2181 2200 2201 2220 2221 2001 2341 2321 2340 2440 2441 2460 2461 2480 2481 2500 2501 2520 2521 2540 2541 2560 2561 2580 2581 2600 739 838 839 938 2360 2361 2380 2381 2400 2401 2420 2421 Ethics 1205 – 1220 Muscle Cells 1221 – 1240 1291 1300 3001 3020 Neural Cell 1241 – 1300 1290 1281 Other 3040 3021 1271 1280 433 532 533 632 832 833 932 Neural Cell 20' Chromatin in 3041 3060 1270 1261 Stem Cells 3080 3061 Posters 2001 – 2600 1251 1260 331 430 431 530 531 630 63 733 831 930 931 1030 20' 3081 3100 1250 1241 3120 3101 Neural Cells (cont.) 2001 – 2064 1240 329 428 629 728 729 830 829 928 929 1028 1231 Muscle Cells 20' 20' 20' 20' 3121 3140 Cancer Cells 2065 – 2150 1230 1221 iPS Cells 3160 3141 427 526 527 626 Germline Cells 2151 – 2170 1211 1220 Ethics 325 724 824 825 924 925 1024 3161 3180 Technologies for 20' 20' 20' 1210 1201 50' 3200 3181 Stem Cell Research 2171 – 2274 Regeneration Epithelial Cells 1191 1200 323 422 623 722 723 823 922 923 1022 3201 3220 1190 1181 Eye or Retinal Cells 20' Tissue Engineering 2275 – 2320 (not skin) Mechanisms 3240 3221 1171 1180 820 3241 20' 20' 20' 720 721 821 920 20' 1020 3260 Totipotency/Early 1170 1161 Intestinal/Gut Cells Reprogramming 3280 3261 Embryo Cells 2321 – 2330 1151 1160 319 418 619 718 719 818 819 918 919 1018 3281 3300 Intern Hematopoietic Stem Cells 2331 – 2416 1150 1141 Epidermal Cells 60' Embryonic Stem Cells 3320 3301 Clinical Application Mesenchymal Cell Endothelial Cells/ Lineage Analysis 2417 – 2443 1131 1140 715 814 815 914 3321 3340 1130 Hemangioblasts 1121 3360 3341 Mesenchymal Stem 1111 1120 313 413 712 713 812 813 912 1012 3361 3380 1110 1101 20' Cell Differentiation 2444 – 2544 30' 30' 3400 3381 311 410 710 711 811 910 1091 1100 20' 20' 3401 3420 Pre-Clinical and Clinical 1090 1081 Cardiac Cells 20' 3440 3421 Applications of 309 408 611 708 709 810 809 908 909 1008 1071 1080 20' 20' 20' Embryonic Stem Cell 3441 3460 Mesenchymal Cells 2545 – 2600 1070 1061 Di erentiation 3480 3461 407 506 507 EXH Lung Cells SER Posters 3001 – 3600 1051 1060 305 404 504 605 704 705 804 805 1004 3481 3500 20' 20' 20' 1050 1041 3520 3501 Other 3001 – 3048 1031 1040 30Liver3 Cells 403 502 503 603 702 703 802 803 Embr902 yonic90 3Stem1002 Cell 20' 20' 3521 3540 1030 1021 Pluripotency 3560 3541 Chromatin in Stem Cells 3049 – 3072 1011 1020 30Pancreatic1 400 Cells401 500 501 600 601 700 701 800 801 900 901 1000 3561 3580 iPS Cells 3073 – 3222 1010 1001 3600 3581 Main Entrance Regeneration Mechanisms 3223 – 3264 Reprogramming 3265 – 3314 Embryonic Stem Cells Clinical Application 3315 – 3340 Embryonic Stem Cell Differentiation 3341 – 3508 Embryonic Stem Cell Pluripotency 3509 – 3600 MAIN ENTRANCE NO PHOTOGRAPHY OR RECORDINGS PERMITTED IN THE POSTER AREAS 99 ATTEND THE SUMMIT OF CUTTING-EDGE CELL BIOLOGY Present, learn, and network at the ASCB Annual Meeting

DeCeMBer 3–7, 2011 | Den Ver, COlOrADO, USA CUTTING-EDGE CELL BIOLOGY—ALL IN ONE pLACE. The ASCB Annual Meeting provides the opportunity to learn new developments in cell biology, present your research, and network with old friends and new contacts. Pick from focused Symposia, Minisymposia, posters, Keynote Address, and more. Visit the exhibits to find tools and resources to help your research. KEYNOTE ADDrESS • Molecules and Systems: Our Quest for a Physiology of the Cell , Harvard Medical School

SYMpOSIA, MINISYMpOSIA, AND MOrE… • Molecular Mechanisms • Function of Multi-Molecular Machines • Cellular networks and Information Processing • Self-Organization of Cellular Structures The science of life, the life of science • Complex Cellular Functions: linking networks and Structures • Mechanisms of Multicellular Functions • Design Principles of Cells and Tissues • Over 30 Minisymposia—Stem Cells and Pluripotency, Cancer Cell Biology and more… register today and take advantage of discounted registration fees until October 3, 2011. For complete details on abstracts, deadlines, special rates, and more, go to:

100 www.ascb.org/meetings www.isscr.org Final Program

Author Index

A C E H Ables, Elizabeth 70 Camargo, Fernando 79 Eggan, Kevin 86 Hammersmith, Katy A. 64 Ader, Marius 68 Candi, Aurélie 88 Elefanty, Andrew G. 80, 84 Handin, Robert I. 75 Agarwal, Suneet 90 Carpenedo, Richard L. 64 Elliott, David 84 Han, Dong-Wook 60 Aifantis, Iannis 63 Casini, Simona 84 Ellis, James 62 Hanjaya-Putra, Donny 65 Alan, Davidson 75 Chen, Antonia 81 Escobar, David 76 Han, Songyan 81 Altschuler, Gabriel 92 Chen, Chien-Wen 85 Essers, Marieke A.G. 74 Harris, James M. 74 Arauzo-Bravo, Marcos 60 Chen, Chun-Chun 75 Ezekowitz, Alan 87 Harris, Lauren J. 74 Atsma, Douwe 84 Chen, Ginny I. 69 Ha, Vi Luan 80 Aviv, Tzvi 66 Chien, Kenneth R. 83 F Heffner, Garrett C. 87 Avruch, Joseph 79 Chon, Andy T. 80 Hegab, Ahmed E. 80 Aznar Benitah, Salvador 68 Chung, Julia 86 Fan, Guoping 89 He, Shenghui 81 Clarke, Geoff 64 Fazzina, Raffaella 90 He, Xi 77 Feldman, Ricardo 84 B Clarke, Ian 66 Hide, Winston 92 Clarke, Michael F. 71 Festa, Eric 79 Hiramatsu, Kunihiko 65 Bai, Lixia 78 Clevers, Hans C. 62 Filomeno, Paola 85 Hochedlinger, Konrad 62, 89 Baptista, Ricardo 64 Coffey, Peter 86 Fischer, Anja 77 Hochgeschwender, Ute 75 Bear, Christine 62 Conboy, Irina M. 82 Fluri, David 64 Holtzinger, Audrey 80 Behbehani, Greg K. 69 Conboy, Michael 82 Foley, Amy 91 Horowitz, Mark 79 Belmonte, Juan Carlos Izpisua 88 Corbel, Stephane 67 Forsthoefel, David 76 Horsley, Valerie 79 Bendall, Sean C. 69 Corneo, Barbara 80 Fretz, Jackie 79 Houghton, Carol 78 Ben-David, Uri 88 Corselli, Mirko 85 Freund, Christian 84 Howard, Jason T. 75 Benvenisty, Nissim 88 Cortes, Mauricio 74 Fuchs, Elaine 77 Hsu, Hwei-Jan 70 Beronja, Slobodan 77 Cox, Andrew G. 74 Fu, Jidong 91 Huang, Kevin 89 Berry, Ryan 79 Crisan, Mihaela 85 Fung, Ho-Lim 88 Huang, Yu 91 Bezzina, Connie 84 Curak, Jasna 69 Huang, Yun 90 Bhutto, Imran 84 Curtis, Daniel 86 G Huard, Johnny 85 Billia, Filio 85 Cutting, Claire C. 74 Hukriede, Neil 75 Gadue, Paul 81 Bisht, Bharti 80 Czechowicz, Agnieszka 74 Huttner, Wieland B. 78 Blanchard, Joel 86 Gage, Fred H. 90 Blanpain, Cédric 88 Gan, Olga I. 88 Blasco, Maria A. 87 D Gao, Xueqin 85 I Blau, Helen M. 67, 91 Dai, Rui 75 Garnaas, Maija K. 74 Ichida, Justin K. 86 Blelloch, Robert 60 Daley, George Q. 59, 80, 87, 90 Gascard, Philippe 70 Ince, Tan 92 Bodenmiller, Bernd 69 Dambrot, Cheryl 84 Gerecht, Sharon 65 Ivanova, Natalia 63 Bose, Vivek 65 Darmawan, Daphne O. 80 Gilbert, Jennifer L. 80 Izpisua Belmonte, Juan Carlos 90 Bouchez, Laure 86 Davies, John E. 85 Gingras, Anne-Claude 69 Giorgetti, Alessandra 90 Bouwmeester, Tewis 86 Davis, Richard 84 J Braam, Stefan 84 Dayan, Victor 85 Glosson, Nicole 73 Bratt-Leal, Andres M. 64 de Groh, Eric 75 Goderie, Susan K. 66 Jaenisch, Rudolf 73 Breuer, Christopher K. 83 De Luca, Michele 83 Goessling, Wolfram 74 James, Noelle 76 Bronner, Marianne 61 Dick, John E. 71, 88 Gomperts, Brigitte N. 80 Jankowska, Anna M. 90 Brummelkamp, Thijn 79 Diep, Cuong 75 Gonska, Tanja 62 Jarvis, Erich D. 75 Buckley, Shannon M. 63 di Fiore, Pier Paolo 70 Goodell, Margaret A. 82 Jeong, Jaemin 82 Burdick, Jason A. 65 Ding, Sheng 84 Gore, Athurva 88 Johnston, Sean 92 Buzina, Alla 88 Dirks, Peter 66 Gouon-Evans, Valerie 81 Djaballah, Hakim 87 Graf, Thomas 72 Dovey, Michael C. 74 Green, Michael D. 81 Drane, Pascal 89 Gurdon, John 72 Drummond-Barbosa, Daniela 70 d’Souza, Sunita L. 81 Dumont, Nancy 70 Dziedzic, Noelle 81 Dzierzak, Elaine 61 101 ISSCR 9th Annual Meeting www.isscr.org

Author Index

K Liu, Yifei 89 Notta, Faiyaz 71 Roy, Soma 70 Lujan, Ernesto 69 Novatchkova, Maria 77 Rubin, Lee 86 Kalatskaya, Irina 88 Lu, Rong 74 Rudnicki, Michael A. 61 Kallin, Eric 72 Lutty, Gerard 84 Rudnicki, Stewart 92 Kaplan, Mark H. 73 O Lutz, Raphael 74 Ruffner, Heinz 86 Keating, Armand 85 Ogawa, Shinichiro 80 Ruiz, Sergio 88 Keller, Gordon 80, 81 Okada, Masaho 85 Kemp, Richard 78 M Ooi, Aik T. 80 Kepple, Kirsten L. 64 Maciejewski, Jaroslaw P. 90 Orkin, Stuart H. 90, 92 S Kern, Andrea 74 Ma, Dongdong 75 Oron, Efrat, 63 Sabine, Charles 84 Ketela, Troy 66, 88 Mak, Anthony B. 69 Otani, Hidetatsu 65 Sacco, Alessandra 91 Khetan, Sudhir 65 Malkoski, Stephen P. 80 Sage, Julien 67 Kiessling, Laura L. 69 Malone, Colin 59 P Saito, Megumu 67 Kim, Carla F. 79 Marchetto, Maria C.N 90 Salathe, Adrian 86 Kim, Hyesoo 87 Markov, Aleksej 91 Pajcini, Kosta 67 Sarangi, Farida 80 Kim, Injune 81 Mayshar, Yoav 88 Paliwal, Preeti 82 Sargent, Carolyn Y. 64 Kim, Jeehee 89 Mazitschek, Ralph 69 Park, Amanda 76 Schaniel, Christoph 81 Kim, Jeong-Beom 60 McDevitt, Todd C. 64 Park, In-Hyun 80 Schebesta, Michael 86 Kim, Suel-Kee 67 McKay, Ronald D. 67 Park, Tea Soon 84 Schlegelmilch, Karin 79 Kinney, Melissa A. 64 Meinhardt, Andrea 68 Pasceri, Peter 62 Schmidt, Barbara 79 Kinzel, Bernd 86 Meissner, Alexander 86 Passier, Robert 84 Schöler, Hans 60 Kirak, Oktay 79 Micallef, Suzanne J. 80 Paw, Barry H. 74 Seita, Jun 74 Kittanakom, Saranya 69 Miller, Diana 84 Peault, Bruno 85 Serluca, Fabrizio 86 Klim, Joseph R. 69 Miller, Freda 67 Pei, Duanqing 63 Shamu, Caroline 92 Knoblich, Juergen A. 77 Milyavsky, Michael 88 Pellegrini, Graziella 83 Shen, Qin 66 Koh, Kian Peng 90 Moffat, Jason 66, 69, 88 Pellegrini, Matteo 80, 89 Shen, Yu-I 65 Kokovay, Erzsi 66 Mohseni, Morvarid 79 Perry, John 77 Silva, Haroldo 82 Ko, Myung Gon 90 Morrison, Sean J. 81 Petrocca, Fabio 92 Simonds, Erin F. 69 Kozar, Sarah 78 Mostoslavsky, Gustavo 90 Plevritis, Sylvia K. 69 Singhal, Nishant 60 Krasnow, Mark 90 Mourkioti, Foteini 67, 91 Porayette, Prashanth 73 Singh, Sean 86 Kriegstein, Arnold 66 Mueller, Matthias 86 Proto, Jonathan 85 Snoeck, Hans-Willem E. 81 Kuzmichev, Andrei 67 Mullighan, Charles 71 Pruszak, Jan 79 Sommer, Cesar A. 90 Mummery, Christine L. 84 Son, Esther Y. 86 L Muotri, Alysson R. 66 Q Sotiropoulou, Panagiotta 88 Mu, Yangling 90 Speck, Nancy 61 Labow, Mark 86 Qian, Li 91 Srivastava, Deepak 91 Lacoste, Arnaud 86 Qiu, Peng 69 Stadtfeld, Matthias 89 LaFever, Leesa 70 N Stagljar, Igor 69 Lam, Kelvin 86 Nagy, Andras 72 R Stainier, Didier 92 Langer, Robert 59 Nakahata, Tatsutoshi 67 Rafuse, Victor 86 Stanley, Edouard G. 80, 84 Latario, Brian 86 Neel, Benjamin G. 69 Rama, Paolo 83 Stary, Joel 76 Le Douarin, Nicole M. 93 Nelson, Brynna 63 Ramirez, Christina 87 Stec, Agnieszka 68 Lee, Gabsang 87 Neumüller, Klaus G. 77 Rangan, Prashanth 59 Stein, Lincoln 88 Lehmann, Ruth 59 Neumüller, Ralph A. 77 Rao, Anjana 90 Stewart, Jocelyn M. 69 Lemischka, Ihor R.2 81 Newmark, Phillip A. 76 Rao, Sridhar 90 Strikoudis, Alexandros 63 Le, Thuc 89 Ng, Elizabeth 84 Ratjen, Felix 62 Studer, Lorenz 87 Lieberman, Judi 92 Nickerson, Derek W. 80 Reya, Tannishtha 73 Sugimura, Ryohichi 77 Li, Lingyin 69 Niesche, Marco 68 Richardson, Andrea 92 Surani, Azim 59 Li, Linheng 77 Niklason, Laura E. 65 Richter, Constance 77 Li, Mo 90 Nishikawa, Shin-Ichi 61 Robin, Catherine 61 Li, Xueling 80 Nishimura, Emi 68 Rodeheffer, Matthew 79 Li, Zhe 88 Nolan, Garry P. 69 Rohrschneider, Larry R. 78 Lim, Megan S. 81 North, Trista E. 74 Rossant, Janet 59, 62 Linderman, Michael D. 69 Nostro, M. Cristina 80, 81 Rossello, Ricardo A. 75 102 www.isscr.org Final Program

Author Index

T W Y Takahashi, Kazutoshi 67 Wagers, Amy 82 Yabuuchi, Akiko 90 Tamplin, Owen J. 76 Wainger, Brian 86 Yadlapalli, Swathi 77 Tanaka, Elly 68 Wang, Jenny 66 Yamanaka, Shinya 67, 71 Tanaka, Takayuki 67 Wang, Xiaoqun 66 Yamashita, Yukiko 77 Tan, Mynn 92 Wang, Xing-Hua 85 Yannarelli, Gustavo 85 Tapia, Natalia 60 Wang, Zheng 63 Yee, Jane 65 Temple, Sally 66 Ward, Dorien 84 Yelon, Debbie 75 Tertoolen, Leon 84 Ward, Ryan 66 Yoder, Mervin C. 73 Thompson, Tadeo O. 62 Waters, Forrest 76 Yokomizo, Tomomasa 61 Tiburcy, Malte 91 Watt, Fiona M. 78 Yoshida, Shosei 60 Tlsty, Thea D. 70 Weissman, Irving L. 59, 74 Yoshikawa, Hideki 65 Tobita, Kimimasa 85 Wenzel, Pamela 87 Yoshimoto, Momoko 73 Toma, Jeremy 86 Wernig, Marius 69 Young, Richard A. 92 Trumpp, Andreas 74 Wilde, Arthur 84 Yousef, Hanadie 82 Tumaki, Noriyuki 65 Williams, Scott E. 77 Yu, Diana 90 Winton, Doug 78 Yu, Juehua 89 U Wong, Amy P. 62 Woolf, Clifford 86 Z Ungrin, Mark 64 Wrighton, Paul J. 69 Wu, Guangming 60 Zambidis, Elias 84 V Wurzer, Stephan 74 Zamparini, Andrea 59 Wu, Tao 89 Zandstra, Peter W. 64 van Delft, Mark F. 88 Zhang, Kelvin X. 80 van den Berg, Cathelijne 84 Zhang, Kun 88 Vander Heiden, Matthew G. 74 X Zhang, Yi 89 Vasioukhin, Valera 79 Xiao, Andrew 89 Zhou, Dawang 79 Venkatraman, Aparna 77 Zhu, Yu 68 Zimmermann, Wolfram H. 91 Zon, Leonard I. 76, 82 Zunder, Eli R. 69

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11th Annual Meeting June 12 – 15, 2013 Boston Convention and Exhibition Center (BCEC) Boston, MA USA www.isscr.org

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