Efficient differentiation of hepatocytes from human embryonic stem cells exhibiting markers recapitulating liver development in vivo
Efficient differentiation of hepatocytes from human embryonic stem cells exhibiting markers recapitulating liver development in vivo
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DOI：
10.1634/stemcells.2007-0718
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发表时间：
2008-04-01
期刊：
影响因子：
5.2
通讯作者：
Cui, Wei
中科院分区：
文献类型：
--
作者：
Hay, David C.;Zhao, Debiao;Cui, Wei
在体外分化人胚胎干细胞（hESC）以提供用于生物医学研究、药物发现和肝病治疗的无限的人肝细胞来源的潜力具有很大的希望。在这里，我们描述了一个三阶段的过程中，高效和可重复的分化hESC肝细胞的启动hESC向定形内胚层与激活素A和丁酸钠，然后进一步分化为肝细胞与二甲亚砜，然后成熟与肝细胞生长因子和制瘤素M。我们已经证明，在这个过程中的hESC分化重演肝脏发育在体内：初始分化后，hESC瞬时表达的原条中内胚层的特征性标志物，然后转向的标志物的定形内胚层;随着进一步分化，表达肝细胞祖细胞标志物和成熟肝细胞标志物依次出现。此外，我们已经提供了证据表明hESC衍生的肝细胞能够执行一系列肝细胞功能：糖原的储存以及血浆蛋白的产生和分泌。更重要的是，hESC衍生的肝细胞表达细胞色素P450同工酶的几个成员，并且这些P450同工酶能够将底物转化为代谢物并响应化学刺激。我们的研究结果提供了证据，hESC可以在体外有效地分化为功能性肝细胞，这可能是有用的体外系统的毒性筛选药物发现。
The potential to differentiate human embryonic stem cells (hESCs) in vitro to provide an unlimited source of human hepatocytes for use in biomedical research, drug discovery, and the treatment of liver diseases holds great promise. Here we describe a three-stage process for the efficient and reproducible differentiation of hESCs to hepatocytes by priming hESCs towards definitive endoderm with activin A and sodium butyrate prior to further differentiation to hepatocytes with dimethyl sulfoxide, followed by maturation with hepatocyte growth factor and oncostatin M. We have demonstrated that differentiation of hESCs in this process recapitulates liver development in vivo: following initial differentiation, hESCs transiently express characteristic markers of the primitive streak mesendoderm before turning to the markers of the definitive endoderm; with further differentiation, expression of hepatocyte progenitor cell markers and mature hepatocyte markers emerged sequentially. Furthermore, we have provided evidence that the hESC-derived hepatocytes are able to carry out a range of hepatocyte functions: storage of glycogen, and generation and secretion of plasma proteins. More importantly, the hESC-derived hepatocytes express several members of cytochrome P450 isozymes, and these P450 isozymes are capable of converting the substrates to metabolites and respond to the chemical stimulation. Our results have provided evidence that hESCs can be differentiated efficiently in vitro to functional hepatocytes, which may be useful as an in vitro system for toxicity screening in drug discovery.