Low-molecular-weight inhibitors of cell differentiation enable efficient growth of mouse iPS cells under feeder-free conditions
Low-molecular-weight inhibitors of cell differentiation enable efficient growth of mouse iPS cells under feeder-free conditions
复制标题
低分子量细胞分化抑制剂使小鼠 iPS 细胞在无饲养层条件下高效生长
DOI：
10.1007/s10616-013-9686-8
复制
发表时间：
2014
期刊：
影响因子：
2.2
通讯作者：
Fukuda T
中科院分区：
文献类型：
--
作者：
Donai K;Inagaki A;So KH;Kuroda K;Sone H;Kobayashi M;Nishimori K;Fukuda T
胚胎干细胞和诱导多能干细胞（iPS）通常维持在来源于小鼠胚胎成纤维细胞（MEF）的饲养细胞上。近年来，iPS细胞在无血清和无饲养层条件下的细胞培养在克服临床应用的生物安全性问题方面越来越受到关注。在这项研究中，我们报告了使用多种小分子抑制剂（即，CHIR 99021、PD 0325901和Thiazovivin）在化学限定和无血清条件下有效培养小鼠iPS细胞而不需要饲养细胞。在这种条件下，我们显示小鼠iPS细胞表达Nanog、Oct 3/4和SSEA-1多能标记物，表明培养条件被优化以维持iPS细胞的多能状态。没有这些小分子抑制剂，小鼠iPS细胞需要适应期才能开始稳定的细胞增殖。这些抑制剂的应用为细胞适应性培养提供了捷径。本研究将有助于在无MEF来源的饲养细胞的情况下有效地建立小鼠iPS细胞系。
Embryonic stem cells and induced pluripotent stem (iPS) cells are usually maintained on feeder cells derived from mouse embryonic fibroblasts (MEFs). In recent years, the cell culture of iPS cells under serum- and feeder-free conditions is gaining attention in overcoming the biosafety issues for clinical applications. In this study, we report on the use of multiple small-molecular inhibitors (i.e., CHIR99021, PD0325901, and Thiazovivin) to efficiently cultivate mouse iPS cells without feeder cells in a chemically-defined and serum-free condition. In this condition, we showed that mouse iPS cells are expressing the Nanog, Oct3/4, and SSEA-1 pluripotent markers, indicating that the culture condition is optimized to maintain the pluripotent status of iPS cells. Without these small-molecular inhibitors, mouse iPS cells required the adaptation period to start the stable cell proliferation. The application of these inhibitors enabled us the shortcut culture method for the cellular adaptation. This study will be useful to efficiently establish mouse iPS cell lines without MEF-derived feeder cells.