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Maintenance of Human Embryonic Stem Cells in Animal Serum- and Feeder Layer-Free Culture Conditions

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Human Embryonic Stem Cell Protocols

Part of the book series: Methods In Molecular Biology ((MIMB,volume 331))

Abstract

The availability of human embryonic stem cells (hESCs) reflects their outstanding potential for research areas such as human developmental biology, teratology, and cellbased therapies. To allow their continuous growth as undifferentiated cells, isolation and culturing were traditionally conducted on mouse embryonic fibroblast feeder layers, using medium supplemented with fetal bovine serum. However, these conditions allow possible exposure of the cells to animal pathogens. Because both research and future clinical application require an animal-free and well-defined culture system for hESCs, these conventional conditions would prevent the use of hESCs in human therapy. This chapter describes optional culture conditions based on either animal-free or feeder-free culture methods for hESCs.

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References

  1. Thomson J. A., Itskovitz-Eldor J., Shapiro S. S., et al. (1998) Embryonic stem cell lines derived from human blastocysts. Science 282, 1145–1147.

    Article  PubMed  CAS  Google Scholar 

  2. Richards M., Fong C. Y., Chan W. K., Wong P. C., and Bongso A. (2002) Human feeders support prolonged undifferentiated growth of human inner cell masses and embryonic stem cells. Nat. Biotechnol. 20, 933–936.

    Article  PubMed  CAS  Google Scholar 

  3. Amit M., Carpenter M. K., Inokuma M. S., et al. (2000) Clonally derived human embryonic stem cell lines maintain pluripotency and proliferative potential for prolonged periods of culture. Dev. Biol. 227, 271–27

    Article  PubMed  CAS  Google Scholar 

  4. Amit M., Margulets V., Segev H., et al. (2003). Human feeder layers for human embryonic stem cells. Biol. Reprod. 68, 2150–2156.

    Article  PubMed  CAS  Google Scholar 

  5. Hovatta O., Mikkola M., Gertow K., et al. (2003). A culture system using human foreskin fibroblasts as feeder cells allows production of human embryonic stem cells. Hum. Reprod. 18, 1404–1409.

    Article  PubMed  Google Scholar 

  6. Cheng L., Hammond H., Ye Z., Zhan X., and Dravid G. (2003). Human adult marrow cells support prolonged expansion of human embryonic stem cells in culture. Stem Cells 21, 131–142.

    Article  PubMed  CAS  Google Scholar 

  7. Xu C., Inokuma M. S., Denham J., et al. (2001) Feeder-free growth of undifferentiated human embryonic stem cells. Nat. Biotechnol. 19, 971–974.

    Article  PubMed  CAS  Google Scholar 

  8. Amit M., Shariki C., Margulets V., and Itskovitz-Eldor J. (2004). Feeder and serum free culture system for human embryonic stem cells. Biol. Reprod. 70, 837–845.

    Article  PubMed  CAS  Google Scholar 

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© 2006 Humana Press Inc.

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Amit, M., Itskovitz-Eldor, J. (2006). Maintenance of Human Embryonic Stem Cells in Animal Serum- and Feeder Layer-Free Culture Conditions. In: Turksen, K. (eds) Human Embryonic Stem Cell Protocols. Methods In Molecular Biology, vol 331. Humana Press. https://doi.org/10.1385/1-59745-046-4:105

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  • DOI: https://doi.org/10.1385/1-59745-046-4:105

  • Publisher Name: Humana Press

  • Print ISBN: 978-1-58829-497-5

  • Online ISBN: 978-1-59745-046-1

  • eBook Packages: Springer Protocols

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