A ROCK inhibitor permits survival of dissociated human embryonic stem cells
A ROCK inhibitor permits survival of dissociated human embryonic stem cells
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DOI：
10.1038/nbt1310
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发表时间：
2007-06-01
影响因子：
46.9
通讯作者：
Sasai, Yoshiki
中科院分区：
文献类型：
--
作者：
Watanabe, Kiichi;Ueno, Morio;Sasai, Yoshiki
人类胚胎干细胞(HES)在细胞分离后存活率低是研究的障碍，阻碍了亚克隆等操作。在这里，我们显示了选择性的Rho相关激酶(ROCK)抑制剂(1，2)，Y-27632，显著地减少了解离诱导的细胞凋亡，提高了克隆效率(从接近1%提高到接近27%)，并促进了基因转移后的亚克隆。此外，用Y-27632处理的分离的HES细胞即使在无血清悬浮培养(3)中也能防止凋亡，并形成漂浮的聚集体。我们证明，Y-27632的保护能力使SFEB培养的HES细胞存活并分化为BF1(+)皮质和基础端脑前体细胞，SFEB培养的小鼠ES细胞也是如此。
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.