Letter abstract
Nature Biotechnology 25, 681 - 686 (2007)
Published online: 27 May 2007 | doi:10.1038/nbt1310
A ROCK inhibitor permits survival of dissociated human embryonic stem cells
Kiichi Watanabe1,5, Morio Ueno1,3, Daisuke Kamiya1, Ayaka Nishiyama1, Michiru Matsumura1, Takafumi Wataya1,4, Jun B Takahashi4, Satomi Nishikawa2, Shin-ichi Nishikawa2, Keiko Muguruma1 & Yoshiki Sasai1
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) inhibitor1, 2, Y-27632, to hES cells markedly diminishes dissociation-induced apoptosis, increases cloning efficiency (from 
1% to 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) culture3 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.
- Organogenesis and Neurogenesis Group, Center for Developmental Biology, RIKEN, Kobe 650-0047, Japan.
- Stem Cell Biology Group, Center for Developmental Biology, RIKEN, Kobe 650-0047, Japan.
- Department of Ophthalmology, National Center for Geriatrics and Gerontology, Obu 474-8511, Japan.
- Department of Neurosurgery and Department of Neurology, Kyoto University Graduate School of Medicine, Kyoto 606-8507, Japan.
- Present address: Division of Biology 216-76, California Institute of Technology, Pasadena, California 91125, USA.
Correspondence to: Yoshiki Sasai1 e-mail: yoshikisasai@cdb.riken.jp
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