Stepwise differentiation of human embryonic stem cells into early endoderm derivatives and their molecular characterization
Stepwise differentiation of human embryonic stem cells into early endoderm derivatives and their molecular characterization
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DOI：
10.1016/j.scr.2011.12.006
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发表时间：
2012-05-01
影响因子：
1.2
通讯作者：
Benvenisty, Nissim
中科院分区：
文献类型：
--
作者：
Kopper, Oded;Benvenisty, Nissim
人类胚胎干细胞具有分化成所有人类细胞类型的潜力，因此具有巨大的治疗前景。分化为胚胎内胚层及其衍生物特别令人感兴趣，因为它可以治愈严重的广泛临床病症，例如糖尿病和肝衰竭。在这项工作中，我们建立了一个独特的实验大纲，可以研究早期人类内胚层发育，并有助于改进和创建新的分化方案。为此，我们从中内胚层细胞开始，并使用 CXCR4 和 PDGFRA 细胞表面标记将它们分离成早期内胚层和中胚层祖细胞。我们对不同谱系进行了分子表征，并证明了 TGF β 途径在定形内胚层起始中的重要性。然后纯化内胚层祖细胞，创建不受其他细胞群影响的内胚层分化生态位。我们跟踪这些细胞在不同时间点的分化，并证明了指示分化为前肠和后肠的基因的上调。令人惊讶的是，在继续培养后，肝基因特征显着下调。这种下调可以通过 FGF2 治疗来挽救，这证明了它在肝细胞维持中的重要性。总之，我们认为分离内胚层祖细胞对于分析其命运至关重要，并且能够鉴定参与其分化和维持的因素。 (C) 2011 Elsevier B.V. 保留所有权利。
Human embryonic stem cells have the potential to differentiate into all human cell types and therefore hold a great therapeutic promise. Differentiation into the embryonic endoderm and its derivatives is of special interest since it can provide a cure for severe widespread clinical conditions such as diabetes and hepatic failure. In this work we established a unique experimental outline that enables the study of early human endoderm development and can help improve and create new differentiation protocols. To this end we started with mesendoderm cells and separated them into early endoderm and mesoderm progenitor cells using CXCR4 and PDGFRA cell surface markers. We molecularly characterized the different lineages, and demonstrated the importance of the TGF beta pathway in definitive endoderm initiation. The endoderm progenitor cells were then purified creating an endodermal differentiation niche that is not affected by other cell populations. We followed the differentiation of these cells at different time points, and demonstrated an up regulation of genes indicative to differentiation into both foregut and hindgut. Surprisingly, upon continued culture, there was significant down regulation of the hepatic gene signature. This down regulation could be rescued with FGF2 treatment demonstrating its importance in hepatic cell maintenance. In conclusion, we suggest that isolating endoderm progenitor cells is crucial for the analysis of their fate, and enables the identification of factors involved in their differentiation and maintenance. (C) 2011 Elsevier B.V. All rights reserved.