A Primitive Growth Factor, NME7AB, Is Sufficient to Induce Stable Naive State Human Pluripotency; Reprogramming in This Novel Growth Factor Confers Superior Differentiation
A Primitive Growth Factor, NME7AB, Is Sufficient to Induce Stable Naive State Human Pluripotency; Reprogramming in This Novel Growth Factor Confers Superior Differentiation
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DOI：
10.1002/stem.2261
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发表时间：
2016-04-01
期刊：
影响因子：
5.2
通讯作者：
Bamdad, Cynthia C.
中科院分区：
文献类型：
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作者：
Carter, M. G.;Smagghe, B. J.;Bamdad, Cynthia C.
科学家们已经培育出在某些方面模仿老鼠幼稚细胞的人类干细胞，但它们对添加几种外在因素的依赖，以及它们发展异常核型的倾向，让人怀疑它们与人类自然出现的幼稚状态的相似性。在这里，我们报道了一种重组的、截短的人NME7，在这里被称为NME7(AB)，在不使用抑制剂、转基因、白血病抑制因子(LIF)、成纤维细胞生长因子2(FGF2)、饲养层细胞或它们的条件培养液的情况下，在人胚胎干细胞中诱导稳定的幼稚状态并诱导多能干细胞。幼稚状态的证据包括雌性来源细胞中第二条X染色体的重新激活，幼稚标记的表达增加和启动状态标记的表达减少，克隆扩展的能力和分化潜力的增加。RNA-seq分析显示，亲本FGF2培养、启动状态细胞和NME7(AB)转化细胞之间存在巨大差异，但与其他通过使用生化抑制剂产生幼稚样干细胞的改变的基因表达模式相似。这里提供的实验，结合我们之前的工作，提出了一个人类干细胞如何调节自我复制的机制模型：由NME7驱动的早期朴素状态，它本身不能限制自我复制，以及后来由NME1调节的朴素状态，当它的多聚体状态从活跃的二聚体转变为不活跃的六聚体时，它限制了自我复制。干细胞2016；34：847-859
Scientists have generated human stem cells that in some respects mimic mouse naive cells, but their dependence on the addition of several extrinsic agents, and their propensity to develop abnormal karyotype calls into question their resemblance to a naturally occurring naive state in humans. Here, we report that a recombinant, truncated human NME7, referred to as NME7(AB) here, induces a stable naive-like state in human embryonic stem cells and induced pluripotent stem cells without the use of inhibitors, transgenes, leukemia inhibitory factor (LIF), fibroblast growth factor 2 (FGF2), feeder cells, or their conditioned media. Evidence of a naive state includes reactivation of the second X chromosome in female source cells, increased expression of naive markers and decreased expression of primed state markers, ability to be clonally expanded and increased differentiation potential. RNA-seq analysis shows vast differences between the parent FGF2 grown, primed state cells, and NME7(AB) converted cells, but similarities to altered gene expression patterns reported by others generating naive-like stem cells via the use of biochemical inhibitors. Experiments presented here, in combination with our previous work, suggest a mechanistic model of how human stem cells regulate self-replication: an early naive state driven by NME7, which cannot itself limit self-replication and a later naive state regulated by NME1, which limits self-replication when its multimerization state shifts from the active dimer to the inactive hexamer. Stem Cells2016;34:847-859