Human embryonic stem cells with biological and epigenetic characteristics similar to those of mouse ESCs
Human embryonic stem cells with biological and epigenetic characteristics similar to those of mouse ESCs
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DOI：
10.1073/pnas.1004584107
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发表时间：
2010-05-18
影响因子：
11.1
通讯作者：
Jaenisch, Rudolf
中科院分区：
文献类型：
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作者：
Hanna, Jacob;Cheng, Albert W.;Jaenisch, Rudolf
人类和小鼠胚胎干细胞（ESC）来源于囊胚期胚胎，但具有非常不同的生物学特性，分子分析表明，迄今为止分离的人类ESC的多能状态与小鼠来源的外胚层干细胞（EpiSC）相对应。在这里，我们将传统人类胚胎干细胞的身份重新连接到一个更不成熟的状态，与多能小鼠胚胎干细胞广泛共享定义特征。这是通过异位诱导Oct 4，Klf 4和Klf 2因子与LIF和糖原合成酶激酶3 β（GSK 3 β）和丝裂原活化蛋白激酶（ERK 1/2）途径的抑制剂组合实现的。毛喉素是一种蛋白激酶A途径激动剂，可诱导Klf 4和Klf 2表达，瞬时替代异位转基因表达的需要。与传统的人ESC相比，这些表观遗传转化的细胞具有与小鼠ESC高度相似的生长特性、X染色体活化状态（XaXa）、基因表达谱和信号传导途径依赖性。最后，相同的生长条件允许衍生具有与小鼠iPS细胞相似性质的人诱导多能干（iPS）细胞。经验证的“幼稚”人类胚胎干细胞的产生将允许对人类先前未定义的多能状态进行分子解剖，并可能为患者特异性疾病相关研究开辟新的机会。
Human and mouse embryonic stem cells (ESCs) are derived from blastocyst-stage embryos but have very different biological properties, and molecular analyses suggest that the pluripotent state of human ESCs isolated so far corresponds to that of mouse-derived epiblast stem cells (EpiSCs). Here we rewire the identity of conventional human ESCs into a more immature state that extensively shares defining features with pluripotent mouse ESCs. This was achieved by ectopic induction of Oct4, Klf4, and Klf2 factors combined with LIF and inhibitors of glycogen synthase kinase 3 beta (GSK3 beta) and mitogen-activated protein kinase (ERK1/2) pathway. Forskolin, a protein kinase A pathway agonist which can induce Klf4 and Klf2 expression, transiently substitutes for the requirement for ectopic transgene expression. In contrast to conventional human ESCs, these epigenetically converted cells have growth properties, an X-chromosome activation state (XaXa), a gene expression profile, and a signaling pathway dependence that are highly similar to those of mouse ESCs. Finally, the same growth conditions allow the derivation of human induced pluripotent stem (iPS) cells with similar properties as mouse iPS cells. The generation of validated "naive" human ESCs will allow the molecular dissection of a previously undefined pluripotent state in humans and may open up new opportunities for patient-specific, disease-relevant research.