title: A ROCK inhibitor permits survival of dissociated human embryonic stem cells.
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.
authors.affiliation: Organogenesis and Neurogenesis Group, Center for Developmental Biology, RIKEN, Kobe 650-0047, Japan.
authors.forename: Kiichi
authors.initials: K
authors.lastname: Watanabe
authors.forename: Morio
authors.initials: M
authors.lastname: Ueno
authors.forename: Daisuke
authors.initials: D
authors.lastname: Kamiya
authors.forename: Ayaka
authors.initials: A
authors.lastname: Nishiyama
authors.forename: Michiru
authors.initials: M
authors.lastname: Matsumura
authors.forename: Takafumi
authors.initials: T
authors.lastname: Wataya
authors.forename: Jun B
authors.initials: JB
authors.lastname: Takahashi
authors.forename: Satomi
authors.initials: S
authors.lastname: Nishikawa
authors.forename: Shin-ichi
authors.initials: S
authors.lastname: Nishikawa
authors.forename: Keiko
authors.initials: K
authors.lastname: Muguruma
authors.forename: Yoshiki
authors.initials: Y
authors.lastname: Sasai
mesh_terms: D000577:Amides / Q000008:administration & dosage*; D000818:Animals; D002454:Cell Differentiation / Q000187:drug effects*; D002470:Cell Survival / Q000187:drug effects*; D002478:Cells, Cultured; D004305:Dose-Response Relationship, Drug; D053595:Embryonic Stem Cells / Q000166:cytology* / Q000378:metabolism*; D006801:Humans; D047908:Intracellular Signaling Peptides and Proteins / Q000037:antagonists & inhibitors* / Q000378:metabolism*; D051379:Mice; D017346:Protein Serine-Threonine Kinases / Q000037:antagonists & inhibitors* / Q000378:metabolism*; D011725:Pyridines / Q000008:administration & dosage*; D054460:rho-Associated Kinases
chemical_list: D000577:Amides; D047908:Intracellular Signaling Peptides and Proteins; D011725:Pyridines; C108830:Y 27632; D017346:Protein Serine-Threonine Kinases; D054460:rho-Associated Kinases