A ROCK inhibitor permits survival of dissociated human embryonic stem cells

  • Nat Biotechnol. 2007 Jun;25(6):681-6. doi: 10.1038/nbt1310.
Kiichi Watanabe  1 ,  Morio Ueno ,  Daisuke Kamiya ,  Ayaka Nishiyama ,  Michiru Matsumura ,  Takafumi Wataya ,  Jun B Takahashi ,  Satomi Nishikawa ,  Shin-ichi Nishikawa ,  Keiko Muguruma ,  Yoshiki Sasai
Affiliations
  • 1. Organogenesis and Neurogenesis Group, Center for Developmental Biology, RIKEN, Kobe 650-0047, Japan.
PMID: 17529971 DOI: 10.1038/nbt1310
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.