Establishment of clinically compliant human embryonic stem cells in an autologous feeder-free system

Tissue Engineering. Part C, Methods
Xin FuTong Cao

Abstract

Applications of human embryonic stem cells (hESCs) are limited by the use of mouse embryonic fibroblasts feeder and animal-derived components during culture. In this study, we demonstrated the potential use of extracellular matrix (ECM) derived from the autologous feeders to support long-term undifferentiated growth of hESCs in xeno-free, serum-free, and feeder-free conditions. Autologous H9 ebF (feeder cells derived from outgrowth of embryoid body [EB] predifferentiated from H9 hESCs) was derived from EBs predifferentiated from H9 hESCs through a direct-plating outgrowth system. The ECM comprising collagen VI, laminin, and fibronectin was extracted from H9 ebF through a freeze-thaw procedure. The autologous ECM together with animal component-free TeSR™2 medium was used to support long-term growth of H1 and H9 hESC lines for up to 20 passages. The maintenance of hESC undifferentiated state by autologous ECM was confirmed by the positive staining of hESC-specific markers (Oct4, SSEA-4, and Tra-1-60) and the expression of pluripotency marker genes (Oct4, Nanog, and Sox2). Flow cytometry further showed that more than 99% of hESCs retained the expression of SSEA-3/4 after long-term culture on autologous ECM. Pluripotency of hESCs o...Continue Reading

References

Apr 1, 1979·The Journal of Cell Biology·K HedmanM Höök
Nov 6, 1998·Science·J A ThomsonJ M Jones
Nov 13, 2001·The Journal of Biological Chemistry·Gloria A Di LulloJames D San Antonio
Nov 27, 2001·Science·E CukiermanK M Yamada
Feb 16, 2002·Methods : a Companion to Methods in Enzymology·K J Livak, T D Schmittgen
Nov 25, 2003·Biology of Reproduction·M AmitJ Itskovitz-Eldor
Jan 20, 2004·Journal of Biomedical Science·Esther R PicacheAxel H Schönthal
Jan 28, 2004·Developmental Dynamics : an Official Publication of the American Association of Anatomists·Elen S RoslerMelissa K Carpenter
Jun 24, 2004·Dermatologic Surgery : Official Publication for American Society for Dermatologic Surgery [et Al.]·Adam M RotundaMichael S Kolodney
Feb 3, 2005·Nature Medicine·Maria J MartinAjit Varki
May 12, 2005·Lancet·Irina KlimanskayaRobert Lanza
Jun 21, 2005·Developmental Dynamics : an Official Publication of the American Association of Anatomists·Eva Sjögren-JanssonPeter S Eriksson
Jun 23, 2005·Biotechnology and Bioengineering·Yan LiRamkumar Mandalam
Jul 14, 2005·Applied Microbiology and Biotechnology·Fernando CoboAngel Concha
Sep 24, 2005·Journal of Cell Science·Ludovic VallierRoger A Pedersen
Oct 22, 2005·Stem Cells·Anne-Catherine FluckigerColette Dehay
Jan 3, 2006·Nature Biotechnology·Tenneille E LudwigJames A Thomson
Feb 9, 2006·Nature Biotechnology·Harry Moore
Apr 6, 2006·Proceedings of the National Academy of Sciences of the United States of America·Jean LuMichael Snyder
Apr 25, 2006·Proceedings of the National Academy of Sciences of the United States of America·Shuyuan YaoSheng Ding
Jun 7, 2006·Biochemical and Biophysical Research Communications·Yanxia LiuHongkui Deng
Sep 15, 2006·Journal of Cellular Physiology·Klaus A BeckerGary S Stein
Oct 31, 2006·Reproduction : the Official Journal of the Society for the Study of Fertility·Heli Skottman, Outi Hovatta
Dec 13, 2006·Methods in Enzymology·Michal Amit, Joseph Itskovitz-Eldor
Apr 27, 2007·Investigative Ophthalmology & Visual Science·Ying GuoSangly P Srinivas
May 24, 2007·Tissue Engineering·Florian SommerAchim Goepferich
Aug 29, 2007·Investigative Ophthalmology & Visual Science·Xiaoqing GuoJeffrey W Ruberti
Aug 30, 2008·Proceedings of the National Academy of Sciences of the United States of America·Miho K FuruePeter W Andrews
May 12, 2009·Journal of Cellular and Molecular Medicine·Wei Seong TohTong Cao

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Citations

Jan 12, 2013·Nature Protocols·Thomas O'LearySusana M Chuva de Sousa Lopes
Dec 15, 2015·Biotechnology Journal·Mohammad M MovahedniaTong Cao
Dec 14, 2012·The Journal of Investigative Dermatology·Fahad K KidwaiTong Cao
Mar 20, 2013·Journal of Orthopaedic Research : Official Publication of the Orthopaedic Research Society·Wei Seong TohMyron Spector
Jan 16, 2013·Journal of Tissue Engineering and Regenerative Medicine·Kai LuTong Cao
Apr 19, 2015·Reproductive Biology and Endocrinology : RB&E·Nina DesaiArsela Gishto
Jan 1, 2013·Cell Regeneration·Jack W LambsheadAndrew L Laslett
Aug 24, 2019·Current Stem Cell Research & Therapy·Tannaz Akbari KolagarSeyed Esmaeil Khoshnam
Sep 27, 2018·Stem Cells International·Firdos Alam KhanSarah Ameen Almofty

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Methods Mentioned

BETA
dissection
flow cytometry
PCR
phase contrast
confocal microscopy

Software Mentioned

ASW
SPSS
FV10
Summit

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