Abstract：
:Realizing the potential of human embryonic stem cells (hESCs) in research and commercial applications requires generic protocols for culture, expansion and genetic modification that function between multiple lines. Here we describe a feeder-free hESC culture protocol that was tested in 13 independent hESC lines derived in five different laboratories. The procedure is based on Matrigel adaptation in mouse embryonic fibroblast conditioned medium (CM) followed by monolayer culture of hESC. When combined, these techniques provide a robust hESC culture platform, suitable for high-efficiency genetic modification via plasmid transfection (using lipofection or electroporation), siRNA knockdown and viral transduction. In contrast to other available protocols, it does not require optimization for individual lines. hESC transiently expressing ectopic genes are obtained within 9 d and stable transgenic lines within 3 weeks.
journal_name
Nat Protocjournal_title
Nature protocolsauthors
Braam SR,Denning C,Matsa E,Young LE,Passier R,Mummery CLdoi
10.1038/nprot.2008.140subject
Has Abstractpub_date
2008-01-01 00:00:00pages
1435-43issue
9eissn
1754-2189issn
1750-2799pii
nprot.2008.140journal_volume
3pub_type
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