Journal
NATURE PROTOCOLS
Volume 6, Issue 7, Pages 1037-1043Publisher
NATURE PUBLISHING GROUP
DOI: 10.1038/nprot.2011.342
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Funding
- University of Michigan
- NSF
- NIH [P20 GM-069985, R01 DE016530]
- NIDCR
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T he culture of human embryonic stem (hES) cells in defined and xenogeneic-free conditions will contribute substantially to future biotechnological and medical applications. To achieve this goal, we developed the first fully defined synthetic polymer coating poly[2-(methacryloyloxy) ethyl dimethyl-(3-sulfopropyl) ammonium hydroxide] (PMEDSAH) that sustains long-term growth of hES cells in different culture media. Here we describe a detailed protocol for the reproducible fabrication of PMEDSAH coating on tissue culture polystyrene dishes, and for the feeder-free culture of hES cells on PMEDSAH coating in defined culture medium. This culture system represents a key step toward the fully defined and xenogeneic-free culture of hES cells.
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