Cellular Biomems
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Characterization of Embryonic Stem Cell-Differentiated Cells As Mesenchymal Stem Cells
The University of Southern Mississippi The Aquila Digital Community Honors Theses Honors College Fall 12-2015 Characterization of Embryonic Stem Cell-Differentiated Cells as Mesenchymal Stem Cells Rachael N. Kuehn University of Southern Mississippi Follow this and additional works at: https://aquila.usm.edu/honors_theses Part of the Cell Biology Commons Recommended Citation Kuehn, Rachael N., "Characterization of Embryonic Stem Cell-Differentiated Cells as Mesenchymal Stem Cells" (2015). Honors Theses. 349. https://aquila.usm.edu/honors_theses/349 This Honors College Thesis is brought to you for free and open access by the Honors College at The Aquila Digital Community. It has been accepted for inclusion in Honors Theses by an authorized administrator of The Aquila Digital Community. For more information, please contact [email protected]. The University of Southern Mississippi Characterization of Embryonic Stem Cell-Differentiated Cells as Mesenchymal Stem Cells by Rachael Nicole Kuehn A Thesis Submitted to the Honors College of The University of Southern Mississippi in Partial Fulfillment of the Requirements for the Degree of Bachelor of Science in the Department of Biological Sciences December 2015 ii Approved by ______________________________ Yanlin Guo, Ph.D., Thesis Adviser Professor of Biological Sciences ______________________________ Shiao Y. Wang, Ph.D., Chair Department of Biological Sciences ______________________________ Ellen Weinauer, Ph.D., Dean Honors College iii ABSTRACT Embryonic stem cells (ESCs), due to their ability to differentiate into different cell types while still maintaining a high proliferation capacity, have been considered as a potential cell source in regenerative medicine. However, current ESC differentiation methods are low yielding and create heterogeneous cell populations. -
Boosting the Cellular Potency of Embryonic Stem Cells by Spliceosome Targeting ✉ Wilfried A
Signal Transduction and Targeted Therapy www.nature.com/sigtrans RESEARCH HIGHLIGHT OPEN Boosting the cellular potency of embryonic stem cells by spliceosome targeting ✉ Wilfried A. Kues1 Signal Transduction and Targeted Therapy (2021) 6:324; https://doi.org/10.1038/s41392-021-00743-9 In recent work published in Cell, Shen et al.1 identified transfected ES cells with short interfering RNAs against different spliceosome inhibition in embryonic stem (ES) cells as a key spliceosome transcripts (the spliceosome consisted of 5 core and mechanism for the transition from pluri- to totipotency. Spliceo- several cofactor subunits, here 14 transcripts were targeted), some inhibition, achieved by RNA interference or the chemical respectively. Transient repression of 10 of the 14 splicing factors inhibitor pladienolide B, may gain widespread relevance to the resulted in ES cells, which maintained the typical colony culture of totipotent ES cells, in vitro differentiation of extra- morphology, however, pluripotent marker genes—Oct4 (Pou5f1), embryonal tissue and organoids, translation to the maintenance of Nanog, Sox2, Zfp42 and others—became down-regulated, at the pluripotent cells of other mammal species, including humans, and same time marker genes of totipotency—particularly Zscan4s and a better molecular understanding of cellular potency in stem cells MERVL—were up-regulated. Zscan4s (Zink finger and SCAN and cancer. domain containing 4) is a transcription factor and MERVL (murine The first successful isolation and maintenance of ES derived endogenous retrovirus L) an endogenous retrovirus with a usually fi 1234567890();,: from the inner cell mass (ICM) of murine blastocyst stages was restricted expression to 2-cell embryos. These results were veri ed described in 1981,2 and since then acted as game changer for by supplementing the culture medium with pladienolide B, a genetic studies in this mammalian model organism. -
A Concise Review on the Classification and Nomenclature of Stem Cells Kök Hücrelerinin S›N›Fland›R›Lmas› Ve Isimlendirilmesine Iliflkin K›Sa Bir Derleme
Review 57 A concise review on the classification and nomenclature of stem cells Kök hücrelerinin s›n›fland›r›lmas› ve isimlendirilmesine iliflkin k›sa bir derleme Alp Can Ankara University Medical School, Department of Histology and Embryology, Ankara, Turkey Abstract Stem cell biology and regenerative medicine is a relatively young field. However, in recent years there has been a tremen- dous interest in stem cells possibly due to their therapeutic potential in disease states. As a classical definition, a stem cell is an undifferentiated cell that can produce daughter cells that can either remain a stem cell in a process called self-renew- al, or commit to a specific cell type via the initiation of a differentiation pathway leading to the production of mature progeny cells. Despite this acknowledged definition, the classification of stem cells has been a perplexing notion that may often raise misconception even among stem cell biologists. Therefore, the aim of this brief review is to give a conceptual approach to classifying the stem cells beginning from the early morula stage totipotent embryonic stem cells to the unipotent tissue-resident adult stem cells, also called tissue-specific stem cells. (Turk J Hematol 2008; 25: 57-9) Key words: Stem cells, embryonic stem cells, tissue-specific stem cells, classification, progeny. Özet Kök hücresi biyolojisi ve onar›msal t›p görece yeni alanlard›r. Buna karfl›n, son y›llarda çeflitli hastal›klarda tedavi amac›yla kullan›labilme potansiyelleri nedeniyle kök hücrelerine ola¤anüstü bir ilgi art›fl› vard›r. Klasik tan›m›yla kök hücresi, kendini yenileme ad› verilen mekanizmayla farkl›laflmadan kendini ço¤altan veya bir dizi farkl›laflma aflamas›ndan geçerek olgun hücrelere dönüflebilen hücrelerdir. -
A Simple Bioreactor-Based Method to Generate Kidney Organoids from Pluripotent Stem Cells
bioRxiv preprint doi: https://doi.org/10.1101/237644; this version posted December 20, 2017. The copyright holder for this preprint (which was not certified by peer review) is the author/funder. All rights reserved. No reuse allowed without permission. A simple bioreactor-based method to generate kidney organoids from pluripotent stem cells Aneta Przepiorskia†, Veronika Sandera†, Tracy Tranb, Jennifer A. Hollywooda, Brie Sorrensona, Jen-Hsing Shiha, Ernst J. Wolvetangc, Andrew P. McMahonb, Teresa M. Holm a, Alan J. Davidson a * a Department of Molecular Medicine & Pathology, University of Auckland, New Zealand. b Department of Stem Cell Biology and Regenerative Medicine, Keck School of Medicine, University of Southern California, USA. c Australian Institute for Bioengineering and Nanotechnology, University of Queensland, Australia. † These authors contributed equally * Corresponding author: [email protected] 1 bioRxiv preprint doi: https://doi.org/10.1101/237644; this version posted December 20, 2017. The copyright holder for this preprint (which was not certified by peer review) is the author/funder. All rights reserved. No reuse allowed without permission. Summary Kidney organoids generated from human pluripotent stem cells have the potential to revolutionize how kidney development and injury are studied. Current protocols are technically complex and suffer from poor reproducibility and high reagent costs restricting scalability. To overcome these issues, we have established a simple, inexpensive and robust method to grow kidney organoids in bulk from human induced pluripotent stem cells. Our organoids develop tubular structures by day (d) 8 and show optimal tissue morphology at d14. A comparison with fetal human kidney suggests that d14 organoid renal structures most closely resemble ‘capillary loop’ stage nephrons. -
Ethical Challenges in Organoid Use
Article Ethical Challenges in Organoid Use Vasiliki Mollaki Hellenic National Bioethics Commission, PC 10674 Athens, Greece; [email protected] Abstract: Organoids hold great promises for numerous applications in biomedicine and biotech- nology. Despite its potential in science, organoid technology poses complex ethical challenges that may hinder any future benefits for patients and society. This study aims to analyze the multifaceted ethical issues raised by organoids and recommend measures that must be taken at various levels to ensure the ethical use and application of this technology. Organoid technology raises several serious ethics issues related to the source of stem cells for organoid creation, informed consent and privacy of cell donors, the moral and legal status of organoids, the potential acquisition of human “characteristics or qualities”, use of gene editing, creation of chimeras, organoid transplantation, commercialization and patentability, issues of equity in the resulting treatments, potential misuse and dual use issues and long-term storage in biobanks. Existing guidelines and regulatory frameworks that are applicable to organoids are also discussed. It is concluded that despite the serious ethical challenges posed by organoid use and biobanking, we have a moral obligation to support organoid research and ensure that we do not lose any of the potential benefits that organoids offer. In this direction, a four-step approach is recommended, which includes existing regulations and guidelines, special regulatory provisions that may be needed, public engagement and continuous monitoring of the rapid advancements in the field. This approach may help maximize the biomedical and social benefits of organoid technology and contribute to future governance models in organoid technology. -
The WNT Target SP5 Negatively Regulates WNT Transcriptional Programs in Human Pluripotent Stem Cells
ARTICLE DOI: 10.1038/s41467-017-01203-1 OPEN The WNT target SP5 negatively regulates WNT transcriptional programs in human pluripotent stem cells Ian J. Huggins1, Tomas Bos1, Olivia Gaylord1, Christina Jessen1, Brianna Lonquich1, Angeline Puranen1, Jenna Richter1, Charlotte Rossdam1, David Brafman2, Terry Gaasterland 3 & Karl Willert 1 The WNT/β-catenin signaling pathway is a prominent player in many developmental pro- cesses, including gastrulation, anterior–posterior axis specification, organ and tissue devel- opment, and homeostasis. Here, we use human pluripotent stem cells (hPSCs) to study the dynamics of the transcriptional response to exogenous activation of the WNT pathway. We describe a mechanism involving the WNT target gene SP5 that leads to termination of the transcriptional program initiated by WNT signaling. Integration of gene expression profiles of wild-type and SP5 mutant cells with genome-wide SP5 binding events reveals that SP5 acts to diminish expression of genes previously activated by the WNT pathway. Furthermore, we show that activation of SP5 by WNT signaling is most robust in cells with developmental potential, such as stem cells. These findings indicate a mechanism by which the develop- mental WNT signaling pathway reins in expression of transcriptional programs. 1 Department of Cellular & Molecular Medicine, University of California San Diego, 9500 Gilman Drive, La Jolla, CA 92093-0695, USA. 2 School of Biological and Health Systems Engineering, Arizona State University, Tempe, AZ 85287, USA. 3 University of California San Diego and Scripps Institution of Oceanography, Scripps Genome Center, 9500 Gilman Drive, La Jolla, CA 92093-0202, USA. Correspondence and requests for materials should be addressed to T.G. -
Electrophysiology Read-Out Tools for Brain-On-Chip Biotechnology
micromachines Review Electrophysiology Read-Out Tools for Brain-on-Chip Biotechnology Csaba Forro 1,2,†, Davide Caron 3,† , Gian Nicola Angotzi 4,†, Vincenzo Gallo 3, Luca Berdondini 4 , Francesca Santoro 1 , Gemma Palazzolo 3,* and Gabriella Panuccio 3,* 1 Tissue Electronics, Fondazione Istituto Italiano di Tecnologia, Largo Barsanti e Matteucci, 53-80125 Naples, Italy; [email protected] (C.F.); [email protected] (F.S.) 2 Department of Chemistry, Stanford University, Stanford, CA 94305, USA 3 Enhanced Regenerative Medicine, Fondazione Istituto Italiano di Tecnologia, Via Morego, 30-16163 Genova, Italy; [email protected] (D.C.); [email protected] (V.G.) 4 Microtechnology for Neuroelectronics, Fondazione Istituto Italiano di Tecnologia, Via Morego, 30-16163 Genova, Italy; [email protected] (G.N.A.); [email protected] (L.B.) * Correspondence: [email protected] (G.P.); [email protected] (G.P.); Tel.: +39-010-2896-884 (G.P.); +39-010-2896-493 (G.P.) † These authors contributed equally to this paper. Abstract: Brain-on-Chip (BoC) biotechnology is emerging as a promising tool for biomedical and pharmaceutical research applied to the neurosciences. At the convergence between lab-on-chip and cell biology, BoC couples in vitro three-dimensional brain-like systems to an engineered microfluidics platform designed to provide an in vivo-like extrinsic microenvironment with the aim of replicating tissue- or organ-level physiological functions. BoC therefore offers the advantage of an in vitro repro- duction of brain structures that is more faithful to the native correlate than what is obtained with conventional cell culture techniques. -
Characterization of the Effects of Bfgf and Gsk3β Inhibitors on Embryoid
University of Arkansas, Fayetteville ScholarWorks@UARK Theses and Dissertations 7-2015 Characterization of the Effects of bFGF and GSK3β Inhibitors on Embryoid Bodies Derived from Human Pluripotent Stem Cells Jonathan Earls University of Arkansas, Fayetteville Follow this and additional works at: http://scholarworks.uark.edu/etd Part of the Molecular Biology Commons Recommended Citation Earls, Jonathan, "Characterization of the Effects of bFGF and GSK3β Inhibitors on Embryoid Bodies Derived from Human Pluripotent Stem Cells" (2015). Theses and Dissertations. 1236. http://scholarworks.uark.edu/etd/1236 This Thesis is brought to you for free and open access by ScholarWorks@UARK. It has been accepted for inclusion in Theses and Dissertations by an authorized administrator of ScholarWorks@UARK. For more information, please contact [email protected], [email protected]. Characterization of the Effects of bFGF and GSK3β Inhibitors on Embryoid Bodies Derived from Human Pluripotent Stem Cells A thesis submitted in partial fulfillment of the requirements for the degree of Master of Science in Biomedical Engineering by Jonathan Earls University of Arkansas Bachelor of Science in Physics, 1998 July 2015 University of Arkansas This thesis is approved for recommendation to the Graduate council. _____________________ Dr. Kaiming Ye Thesis Director _____________________ ______________________ Dr. Sha Jin Dr. Robert Beitle Thesis Co-director: Thesis Committee ABSTRACT Embryoid body (EB) formation is a common first step in many human pluripotent stem cell (hPSC) differentiation protocols. Previous work suggests that EBs are sensitive to growth factor withdrawal if they are derived from hPSCs maintained in feeder independent media such as mTeSR1. To promote cell survival, EBs generated from mTeSR1-adapted hPSCs are sometimes cultured in a medium that contains basic fibroblast growth factor (bFGF), a trophic factor often used in hPSC cultures to maintain self-renewal. -
Cellular Biomems
Chapter 11. Cellular BioMEMS Cellular BioMEMS Academic and Research Staff Professor Joel Voldman Postdoctoral Associates Alison Skelley, Katarina Blagović, Wei Mong Tsang Graduate Students Salil Desai, Lily Kim, Joseph Kovac, Hsu-Yi Lee , Nick Mittal, Katya Puchala, Laralynne Przybyla, Adam Rosenthal, Somponnat Sampattavanich, Brian Taff, Michael Vahey Undergraduate Students Stephanie Cheng, Asiri Ediriwickrema, Stephanie Flavin, Shanette Go, Alice Macdonald, Brianna Petrone, Hari Singhal Technical and Support Staff Chadwick Collins Laboratory for Cellular BioMEMS – Research Themes Our group performs research on BioMEMS, applying microfabrication technology to illuminate biological systems, especially at the cellular level. Specifically, we develop technologies that are used to manipulate cells or make measurements from them. Our research builds upon various disciplines: electrical engineering, microfabrication, bioengineering, surface science, fluid mechanics, mass transport, etc. We take a quantitative approach to designing our technology, using both analytical and numerical modeling to gain fundamental understanding of the technologies that we create. We then take our designs through microfabrication to packaging and testing and to biological assay. Our applications have a strong emphasis on stem cell biology and cell screening. 1. Single-cell manipulation platforms for cell cytometry Sponsors NIH NCRR NSF Graduate Research Fellowship NDSEG Graduate Research Fellowship Singapore-MIT Alliance Project Staff Salil Desai, Nick Mittal, Joseph Kovac, Brian Taff, Brianna Petrone, Hari Singhal Figure 1: A generic single-cell manipulation platform. In such devices individual cells are organized in unique Overview addressable locations for a host of different analyses. As an example format, we show a two-dimensional array of traps. The goal of this research is In functional form, we aim to localize distinct cell populations the development of a series to various portions of the device surface. -
Advances in Adult and Non-Embryonic Stem Cell Research
S. HRG. 108–949 ADVANCES IN ADULT AND NON-EMBRYONIC STEM CELL RESEARCH HEARING BEFORE THE SUBCOMMITTEE ON SCIENCE, TECHNOLOGY, AND SPACE OF THE COMMITTEE ON COMMERCE, SCIENCE, AND TRANSPORTATION UNITED STATES SENATE ONE HUNDRED EIGHTH CONGRESS FIRST SESSION JUNE 12, 2003 Printed for the use of the Committee on Commerce, Science, and Transportation ( U.S. GOVERNMENT PRINTING OFFICE 80–903 PDF WASHINGTON : 2013 For sale by the Superintendent of Documents, U.S. Government Printing Office Internet: bookstore.gpo.gov Phone: toll free (866) 512–1800; DC area (202) 512–1800 Fax: (202) 512–2104 Mail: Stop IDCC, Washington, DC 20402–0001 VerDate Nov 24 2008 06:41 May 20, 2013 Jkt 075679 PO 00000 Frm 00001 Fmt 5011 Sfmt 5011 S:\GPO\DOCS\80903.TXT JACKIE SENATE COMMITTEE ON COMMERCE, SCIENCE, AND TRANSPORTATION ONE HUNDRED EIGHTH CONGRESS FIRST SESSION JOHN MCCAIN, Arizona, Chairman TED STEVENS, Alaska ERNEST F. HOLLINGS, South Carolina, CONRAD BURNS, Montana Ranking TRENT LOTT, Mississippi DANIEL K. INOUYE, Hawaii KAY BAILEY HUTCHISON, Texas JOHN D. ROCKEFELLER IV, West Virginia OLYMPIA J. SNOWE, Maine JOHN F. KERRY, Massachusetts SAM BROWNBACK, Kansas JOHN B. BREAUX, Louisiana GORDON H. SMITH, Oregon BYRON L. DORGAN, North Dakota PETER G. FITZGERALD, Illinois RON WYDEN, Oregon JOHN ENSIGN, Nevada BARBARA BOXER, California GEORGE ALLEN, Virginia BILL NELSON, Florida JOHN E. SUNUNU, New Hampshire MARIA CANTWELL, Washington FRANK R. LAUTENBERG, New Jersey JEANNE BUMPUS, Republican Staff Director and General Counsel ROBERT W. CHAMBERLIN, Republican Chief Counsel KEVIN D. KAYES, Democratic Staff Director and Chief Counsel GREGG ELIAS, Democratic General Counsel SUBCOMMITTEE ON SCIENCE, TECHNOLOGY, AND SPACE SAM BROWNBACK, Kansas, Chairman TED STEVENS, Alaska JOHN B. -
Cell Mechanics in Embryoid Bodies
cells Review Cell Mechanics in Embryoid Bodies Kira Zeevaert 1,2, Mohamed H. Elsafi Mabrouk 1,2 , Wolfgang Wagner 1,2,* and Roman Goetzke 1,2,* 1 Helmholtz-Institute for Biomedical Engineering, Stem Cell Biology and Cellular Engineering, RWTH Aachen University Medical School, 52074 Aachen, Germany; [email protected] (K.Z.); [email protected] (M.H.E.M.) 2 Institute for Biomedical Engineering–Cell Biology, RWTH Aachen University Medical School, 52074 Aachen, Germany * Correspondence: [email protected] (W.W.); [email protected] (R.G.); Tel.: +49-241-80-88611 (W.W.); +49-241-80-80268 (R.G.) Received: 15 September 2020; Accepted: 9 October 2020; Published: 11 October 2020 Abstract: Embryoid bodies (EBs) resemble self-organizing aggregates of pluripotent stem cells that recapitulate some aspects of early embryogenesis. Within few days, the cells undergo a transition from rather homogeneous epithelial-like pluripotent stem cell colonies into a three-dimensional organization of various cell types with multifaceted cell–cell interactions and lumen formation—a process associated with repetitive epithelial-mesenchymal transitions. In the last few years, culture methods have further evolved to better control EB size, growth, cellular composition, and organization—e.g., by the addition of morphogens or different extracellular matrix molecules. There is a growing perception that the mechanical properties, cell mechanics, and cell signaling during EB development are also influenced by physical cues to better guide lineage specification; substrate elasticity and topography are relevant, as well as shear stress and mechanical strain. Epithelial structures outside and inside EBs support the integrity of the cell aggregates and counteract mechanical stress. -
An Introduction to Stem Cell Biology
An Introduction to Stem Cell Biology Michael L. Shelanski, MD,PhD Professor of Pathology and Cell Biology Columbia University Figures adapted from ISSCR. Presentations of Drs. Martin Pera (Monash University), Dr.Susan Kadereit, Children’s Hospital, Boston and Dr. Catherine Verfaillie, University of Minnesota Science 1999, 283: 534-537 PNAS 1999, 96: 14482-14486 Turning Blood into Brain: Cells Bearing Neuronal Antigens Generated in Vitro from Bone Marrow Science 2000, 290:1779-1782 From Marrow to Brain: Expression of Neuronal Phenotypes in Adult Mice Mezey, E., Chandross, K.J., Harta, G., Maki, R.A., McKercher, S.R. Science 2000, 290:1775-1779 Brazelton, T.R., Rossi, F.M., Keshet, G.I., Blau, H.M. Nature 2001, 410:701-705 Nat Med 2000, 11: 1229-1234 Stem Cell FAQs Do you need to get one from an egg? Must you sacrifice an Embryo? What is an ES cell? What about adult stem cells or cord blood stem cells Why can’t this work be done in animals? Are “cures” on the horizon? Will this lead to human cloning – human spare parts factories? Are we going to make a Frankenstein? What is a stem cell? A primitive cell which can either self renew (reproduce itself) or give rise to more specialised cell types The stem cell is the ancestor at the top of the family tree of related cell types. One blood stem cell gives rise to red cells, white cells and platelets Stem Cells Vary in their Developmental capacity A multipotent cell can give rise to several types of mature cell A pluripotent cell can give rise to all types of adult tissue cells plus extraembryonic tissue: cells which support embryonic development A totipotent cell can give rise to a new individual given appropriate maternal support The Fertilized Egg The “Ultimate” Stem Cell – the Newly Fertilized Egg (one Cell) will give rise to all the cells and tissues of the adult animal.