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- Title
A ROCK inhibitor permits survival of dissociated human embryonic stem cells.
- Authors
Watanabe, Kiichi; Ueno, Morio; Kamiya, Daisuke; Nishiyama, Ayaka; Matsumura, Michiru; Wataya, Takafumi; Takahashi, Jun B; Nishikawa, Satomi; Nishikawa, Shin-ichi; Muguruma, Keiko; Sasai, Yoshiki
- Abstract
Poor survival of human embryonic stem (hES) cells after cell dissociation is an obstacle to research, hindering manipulations such as subcloning. Here we show that application of a selective Rho-associated kinase (ROCK) inhibitor, Y-27632, to hES cells markedly diminishes dissociation-induced apoptosis, increases cloning efficiency (from approximately 1% to approximately 27%) and facilitates subcloning after gene transfer. Furthermore, dissociated hES cells treated with Y-27632 are protected from apoptosis even in serum-free suspension (SFEB) culture and form floating aggregates. We demonstrate that the protective ability of Y-27632 enables SFEB-cultured hES cells to survive and differentiate into Bf1(+) cortical and basal telencephalic progenitors, as do SFEB-cultured mouse ES cells.
- Publication
Nature biotechnology, 2007, Vol 25, Issue 6, p681
- ISSN
1087-0156
- Publication type
Journal Article
- DOI
10.1038/nbt1310