Frontiers in Stem Cells and Regeneration Advanced Training Course September 27 - October 3, 2015 MBL • Woods Hole, MA Course Directors: Gerald Schatten, PhD Jennifer Morgan, PhD Pittsburgh Development Center Marine Biological Laboratory Organizing Committee: Diane Carlisle, PhD; Gloria Perez, DVM; Dorianne Mebane MBA All lectures are in Loeb G70 (south), unless otherwise specified. Lab exercises are held in Loeb 306. Meals are in the Swope dining room, 2nd floor. Breakfast: 7-8:30 AM; Lunch: 11:30 AM-1 PM; Dinner: 5-7 PM. Coffee breaks will be served in the Loeb ground floor corridor adjacent to the lecture hall. Sunday, September 27 5:30 PM Dinner, Café Swope (first floor) 6:30 PM Lecture: Introduction to Regeneration Ken Muneoka, Texas A&M 7:15 PM Lecture: Challenges in Stem Cells and Regeneration Jerry Schatten, U. Pittsburgh 8:00 PM Lecture: Bioethics and Stem Cell Research Eli Adashi, Brown 9:00 PM EEO Information and Logistics: Biosafety, Regulatory Restrictions, MOUs, and Team Assignments Jennifer Morgan, MBL Monday, September 28 8:30 AM Lecture: Melanoma Stem Cells – Biology and Implications for Immune Checkpoint Therapy Tobias Schatton, Harvard/ Brigham and Women’s 9:15 AM Lecture: Regeneration and Engineering of the Blood System- a means to treat Cancer and Genetic Disorders Amittha Wickrema, UChicago 10:00 AM Coffee Break 10:15 AM Lecture: Patient Specific Pluripotent Stem Cells Charles Easley, Emory 10:45 AM Role of Somatic Stem Cells in Regenerative Medicine Jerry Shay, U. Texas Southwestern Medical Center 11:30 AM Lab Introduction: IPS Cell Derivation, Culture, and Differentiation Charles Easley 11:45 AM Lab Introduction: Calvin Simerly - mESC derivation Noon Lunch 1:00 PM Lab 1: Instructor: Chas Easley. TA: Alyse Stevens. Students will learn how to generate human iPSCs using Cytotune 2.0 (Sendai virus) and perform the initial steps of reprogramming. Students will also learn how to isolate colonies from reprogrammed lines and then learn how to passage established human iPSCs/ESCs. Lab 2: Instructor: Cal Simerly. TA:Zachary McEachin. Mouse embryonic stem cell (mESC) derivation: Students will do immunosurgery technique on mouse expanded blastocysts to isolate the ICM in order to derive mESCs. 6:00 PM Dinner 7:00 PM PIONEER LECTURE: Epigenetics Hunt Willard; Director of MBL 8:00 PM Course Participants’ Introductions Tuesday, September 29 8:30 AM Lecture: Limb Regeneration David Gardiner, UC Irvine 9:15 AM Lecture: Molecular Basis of Limb Regeneration Randal Voss, U Kentucky 10:00 AM Coffee Break 10:15 AM Lecture and Lab Intro: Stem cells and Regeneration in Planarians David Forsthoefel, U. Illinois 11:15 AM Lab Introduction: David Gardiner and Randall Voss - Salamander regeneration 11:30 AM Special Topics Lunch – with vendor sponsors 1:00 PM Lab 1: Instructors: David Gardiner, Randal Voss. TA: Shambhavi Singh (UC Irvine). Regeneration in Salamanders: induction of ectopic limb regeneration Lab 2: Instructor: David Forsthoefel, U. Illinois Regeneration in Planarians 2 6:00 PM Dinner 7:00 PM Course Participants’ Introductions Wednesday, September 30 8:30 AM Lecture: Picking your favorite - cell identification and selection strategies in human neural stem cell biology Jan Pruszak, U. Freiburg 9:15 AM Lecture: Spinal Cord Regeneration in a Non-Mammalian Model Jen Morgan, MBL 10:00 AM Coffee Break 10:15 AM Lecture: Modeling Alzheimer's disease in vitro Maya Mitalipova, MIT 11:00 AM Lab Introduction: Jan Pruszak - Isolating and Characterizing Neural stem cells using FACS strategy. 11:30 AM Lab Introduction: Jen Morgan - Lamprey model of spinal cord injury and regeneration Noon Lunch 1:00 PM Lab 1: Instructor: Jan Pruszak. TA: David Lee (BD Biosciences), Nisha Raj (Emory). In this laboratory, trainees will learn the techniques for isolating specific PSC-derived subsets of therapeutic and scientific relevance using flow cytometry. Lab 2: Instructor: Jen Morgan. TA: Scott Allen (MBL). Students will learn to quantify behavioral recovery after spinal cord injury and will examine various aspects of nervous tissue regeneration (e.g. axons, neurons, glial cells). 6:00 PM Dinner 7:00 PM Discussion: Stem Cell and Regeneration Research Funding Ravi Ravindranath, NIH 8:00 PM Career Mentoring Discussion Facilitated by Jennifer Morgan (MBL) and other course faculty (*Please bring a copy of your CV) Thursday, October 1 8:30 AM Lecture: Translating Spermatogonial Stem Cell Transplantation to the Clinic 3 Kyle Orwig, U. Pittsburgh 9:00 AM Coffee Break 9:15 AM Lecture: Directed Differentiation and hESC as Developmental Sensors Diane Carlisle, U. Pittsburgh 10:00 AM Lab Intro: Directed differentiation Diane Carlisle, U. Pittsburgh 10:30 AM Lab Intro: Kyle Orwig, U. Pittsburgh 11:00 AM Orwig Lab Prep: Differential separation of cells and apply antibodies for staining Carlisle Lab Prep: Preparing differentiated cells for IF staining Noon Lunch 1:00 PM Lab 1: Instructor: Diane Carlisle. TA: Minnu Suresh. We will use differential immunocytochemistry staining to identify enriched populations in cells after directed differentiation. Lab 2: Instructor: Kyle Orwig. TAs: Meena Sukhwani (U. Pittsburgh), Adetunji Fayomi (U. Pittsburgh). Germ stem cell isolation, identification, and transplantation using a magnetic sorting (MACS) strategy. 6:00 PM Dinner 7:00 PM Lecture: Retinoic Acid and Stem Cells James Monaghan, Northeastern University Malcolm Maden, U. Florida 8:30 PM T-Shirt Night Celebration Friday, October 2 8:30 AM Lecture: Echinoderm models of gut and nerve regeneration Jose Garcia-Arraras, U. Puerto Rico 9:15 AM Lecture: Stem Cells in Lung Development and Renewal Mark Krasnow, Stanford 10:00 AM Coffee Break 10:15 AM Preimplantation Genetic Diagnosis Mark Hughes, Genesis Genetics 11:00 AM Lab Introduction: Jose Garcia-Arraras - Echinoderm regeneration 4 11:15 AM Lab Introduction for ongoing experiments: Kyle Orwig, Charles Easley, David Gardiner, Calvin Simerly 11:30 AM Course Photo Noon Lunch 1:00 PM Lab 1: Instructor: Jose Garcia-Arraras. Echinoderm regeneration Lab 2: Finish all ongoing experiments: Instructor: Kyle Orwig. TA: Meena Sukhwani. Staining of enriched germ cell populations. Instructor: Chas Easley. Analysis of iPSCs using microscopy. Check on your hESC culture. Instructors: David Gardiner, Randall Voss. TA:Kate McCusker. Observe blastema formation on regenerating salamanders. Instructor: Cal Simerly. Check on mESC derivation. 5:00 PM PIONEER LECTURE: Stem Cells and Regeneration Phil Newmark, U. Illinois 6:30 PM Banquet: Café Swope 8:00 PM Presentation of Pioneer Awards and Student Certificates Jennifer Morgan Saturday, Oct 3 Check out of rooms before 10:00 AM Thank you to the following course sponsors who have contributed to the course by loaning equipment and/or donating reagents and supplies: 5 .