A ROCK inhibitor permits survival of dissociated human embryonic stem cells

Kiichi Watanabe, Morio Ueno, Daisuke Kamiya, Ayaka Nishiyama, Michiru Matsumura, Takafumi Wataya, Jun B. Takahashi, Satomi Nishikawa, Shin Ichi Nishikawa, Keiko Muguruma, Yoshiki Sasai

Research output: Contribution to journalArticlepeer-review

1687 Citations (Scopus)


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 ∼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) 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.

Original languageEnglish
Pages (from-to)681-686
Number of pages6
JournalNature Biotechnology
Issue number6
Publication statusPublished - 06-2007
Externally publishedYes

All Science Journal Classification (ASJC) codes

  • Biotechnology
  • Bioengineering
  • Applied Microbiology and Biotechnology
  • Molecular Medicine
  • Biomedical Engineering


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