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| A ROCK inhibitor permits survival of dissociated human embryonic stem cells |
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| Author(s): Watanabe K (Watanabe, Kiichi), Ueno M (Ueno, Morio), Kamiya D (Kamiya, Daisuke), Nishiyama A (Nishiyama, Ayaka), Matsumura M (Matsumura, Michiru), Wataya T (Wataya, Takafumi), Takahashi JB (Takahashi, Jun B.), Nishikawa S (Nishikawa, Satomi), Nishikawa S (Nishikawa, Shin-ichi), Muguruma K (Muguruma, Keiko), Sasai Y (Sasai, Yoshiki) |
| Source: NATURE BIOTECHNOLOGY Volume: 25 Issue: 6 Pages: 681-686 Published: JUN 2007 |
| Times Cited: 106 References: 30 |
| 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(1,2), Y-27632, to hES cells markedly diminishes dissociation- induced apoptosis, increases cloning efficiency (from similar to 1% to similar 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) culture(3) 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. |
| Document Type: Article |
| Language: English |
| Reprint Address: Sasai, Y (reprint author), RIKEN, Ctr Dev Biol, Organogenesis & Neurogenesis Grp, Kobe, Hyogo 6500047 Japan |
Addresses:
1. RIKEN, Ctr Dev Biol, Organogenesis & Neurogenesis Grp, Kobe, Hyogo 6500047 Japan
2. RIKEN, Ctr Dev Biol, Stem Cell Biol Grp, Kobe, Hyogo 6500047 Japan
3. Natl Ctr Geriatr & Gerontol, Dept Ophthalmol, Oulu 4748511, Finland
4. Kyoto Univ, Grad Sch Med, Dept Neurosurg, Kyoto 6068507, Japan
5. Kyoto Univ, Grad Sch Med, Dept Neurol, Kyoto 6068507, Japan |
| Publisher: NATURE PUBLISHING GROUP, 75 VARICK STREET, 9TH FLOOR, NEW YORK, NY 10013-1917 USA |
| Subject Category: Biotechnology & Applied Microbiology |
| IDS Number: 176IF |
| ISSN: 1087-0156 |
| DOI: 10.1038/nbt1310 |
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