Systematic identification of culture conditions for induction and maintenance of naive human pluripotency.
Systematic identification of culture conditions for induction and maintenance of naive human pluripotency.
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系统地识别培养条件，以诱导和维持幼稚的人类多能。
DOI：
10.1016/j.stem.2014.07.002
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发表时间：
2014-10-02
期刊：
影响因子：
23.9
通讯作者：
Jaenisch, Rudolf
中科院分区：
文献类型：
--
作者：
Theunissen, Thorold W.;Powell, Benjamin E.;Wang, Haoyi;Mitalipova, Maya;Faddah, Dina A.;Reddy, Jessica;Fan, Zi Peng;Maetzel, Dorothea;Ganz, Kibibi;Shi, Linyu;Lungjangwa, Tenzin;Imsoonthornruksa, Sumeth;Stelzer, Yonatan;Rangarajan, Sudharshan;D'Alessio, Ana;Zhang, Jianming;Gao, Qing;Dawlaty, Meelad M.;Young, Richard A.;Gray, Nathanael S.;Jaenisch, Rudolf
小鼠和人类的胚胎干细胞（ESC）具有不同的分子和生物学特征，这就提出了一个问题，即人类是否存在早期的“幼稚”多能性状态。在这里，我们采取了一种系统的方法来鉴定支持幼稚人ESC自我更新的小分子，其基于内源性OCT 4远端增强子活性的维持，这是基态多能性的分子特征。迭代化学筛选确定了五种激酶抑制剂的组合，当直接应用于常规人类ESC时，其诱导并维持OCT 4远端增强子活性。这些抑制剂产生人多能细胞，其中与多能性基态相关的转录因子被高度上调并且二价染色质结构域被耗尽。与先前报道的幼稚人类胚胎干细胞的比较表明，我们的条件捕获一个独特的多能状态，在人类中，非常相似的小鼠胚胎干细胞。这项研究提出了一个框架，用于定义幼稚人类多能细胞的培养要求。TALEN介导的用于幼稚人类多能性的报告系统的工程化在不存在转基因的情况下维持幼稚报告活性的化学筛选优化的化学条件捕获人类多能性的独特状态幼稚人类细胞的基因表达与幼稚小鼠细胞的基因表达高度相似通过顺序化学筛选，泰尼森et al.鉴定激酶抑制剂的组合，其诱导并维持人胚胎干细胞中幼稚多能性的限定特征。
Embryonic stem cells (ESCs) of mice and humans have distinct molecular and biological characteristics, raising the question of whether an earlier, “naive” state of pluripotency may exist in humans. Here we took a systematic approach to identify small molecules that support self-renewal of naive human ESCs based on maintenance of endogenous OCT4 distal enhancer activity, a molecular signature of ground state pluripotency. Iterative chemical screening identified a combination of five kinase inhibitors that induces and maintains OCT4 distal enhancer activity when applied directly to conventional human ESCs. These inhibitors generate human pluripotent cells in which transcription factors associated with the ground state of pluripotency are highly upregulated and bivalent chromatin domains are depleted. Comparison with previously reported naive human ESCs indicates that our conditions capture a distinct pluripotent state in humans that closely resembles that of mouse ESCs. This study presents a framework for defining the culture requirements of naive human pluripotent cells. TALEN-mediated engineering of a reporter system for naive human pluripotency Chemical screen for maintenance of naive reporter activity in absence of transgenes Optimized chemical conditions capture a distinct state of human pluripotency Gene expression of naive human cells is highly similar to that of naive mouse cells Through sequential chemical screening, Theunissen et al. identify a combination of kinase inhibitors that induces and maintains defining features of naive pluripotency in human embryonic stem cells.
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