Propagation of Human Embryonic Stem Cells on Human Feeder Cells

  • Mark Richards
  • Ariff Bongso
Part of the Methods In Molecular Biology book series (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.

Key Words

Bulk culture &quote;cut and paste&quote; protocol human adult fibroblasts human embryonic stem cells human fetal fibroblasts nonsupportive feeder supportive feeder 

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Copyright information

© Humana Press Inc. 2006

Authors and Affiliations

  • Mark Richards
    • 1
  • Ariff Bongso
    • 1
  1. 1.Department of Obstetrics and GynecologyNational University Hospital, National University of SingaporeKent RidgeSingapore

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