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Propagation of Human Embryonic Stem Cells on Human Feeder Cells

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Human Embryonic Stem Cell Protocols

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

Abstract

Human embryonic stem (hES) cell lines are usually derived and propagated on inactivated murine embryonic fibroblast (MEF) feeders. The use of MEFs and culture ingredients of animal origin for hES cell support increases the risk of cross-contamination of the hES cells with infectious animal agents from the MEFs and animal-based culture medium. This thus makes such hES cells lines undesirable for clinical application. This chapter describes several protocols used in the propagation of hES cells on human fibroblast feeder cells. Two culture methods, the bulk enzymatic culture protocol and the microdissection “cut and paste” protocol are described. Only certain human fetal and adult fibroblast feeders support hES cell growth. Methods for the characterization of pluripotent undifferentiated hES cells grown on human feeders including cell surface marker staining and real-time polymerase chain reaction are also described.

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Authors and Affiliations

  1. Department of Obstetrics and Gynecology, National University Hospital, National University of Singapore, Kent Ridge, Singapore

    Mark Richards & Ariff Bongso

Authors
  1. Mark Richards
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  2. Ariff Bongso
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Editor information

Editors and Affiliations

  1. Ottawa Health Research Institute, Ottawa, Ontario, Canada

    Kursad Turksen

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© 2006 Humana Press Inc.

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Richards, M., Bongso, A. (2006). Propagation of Human Embryonic Stem Cells on Human Feeder Cells. In: Turksen, K. (eds) Human Embryonic Stem Cell Protocols. Methods In Molecular Biology, vol 331. Humana Press. https://doi.org/10.1385/1-59745-046-4:23

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  • DOI: https://doi.org/10.1385/1-59745-046-4:23

  • 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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