TY - JOUR
T1 - A ROCK inhibitor permits survival of dissociated human embryonic stem cells
AU - Watanabe, Kiichi
AU - Ueno, Morio
AU - Kamiya, Daisuke
AU - Nishiyama, Ayaka
AU - Matsumura, Michiru
AU - Wataya, Takafumi
AU - Takahashi, Jun B.
AU - Nishikawa, Satomi
AU - Nishikawa, Shin Ichi
AU - Muguruma, Keiko
AU - Sasai, Yoshiki
N1 - Funding Information:
We are grateful to S. Narumiya and H. Bito for discussions and invaluable advice about ROCK inhibitors, to M. Hirose for kind advice and help in cell cycle analysis, to H. Niwa, H. Enomoto and N. Love for discussion and technical advice. Y.S. is thankful to Kenzo Sasai and Tetsuro Haraguchi, who passed away while this project was underway, for continuous encouragement. This work was supported by grants-in-aid from Ministry of Education, Culture, Sports, Science and Technology, the Kobe Cluster Project and the Leading Project (Y.S., S.N., J.B.T.). The hES cells (KhES-1, 2 and 3) were a gift from N. Nakatsuji and H. Suemori (Kyoto University) and the drug-selectable plasmid pCAGGS-Venus-Hygro was a gift from H. Niwa.
PY - 2007/6
Y1 - 2007/6
N2 - 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.
AB - 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.
UR - https://www.scopus.com/pages/publications/34250206996
UR - https://www.scopus.com/pages/publications/34250206996#tab=citedBy
U2 - 10.1038/nbt1310
DO - 10.1038/nbt1310
M3 - Article
C2 - 17529971
AN - SCOPUS:34250206996
SN - 1087-0156
VL - 25
SP - 681
EP - 686
JO - Nature Biotechnology
JF - Nature Biotechnology
IS - 6
ER -