Efficient differentiation of functional hepatocytes from human embryonic stem cells
Efficient differentiation of functional hepatocytes from human embryonic stem cells
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DOI：
10.1634/stemcells.2007-1102
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发表时间：
2008-05-01
期刊：
影响因子：
5.2
通讯作者：
Lanza, Robert
中科院分区：
文献类型：
--
作者：
Agarwal, Sadhana;Holton, Katherine L.;Lanza, Robert
人胚胎干细胞（hESC）分化为特定功能细胞类型可以使用模拟体内胚胎发育程序的方法来实现。目前用于从hESC产生肝细胞的方案受到低效分化程序的阻碍，所述分化程序导致低产量和大的细胞异质性。我们在这里报告了一个强大的和高效的过程中产生的高纯度（70%）的肝细胞培养的hESC，平行于顺序的肝发育在体内。由hESC产生高度富集的定形内胚层群体，然后通过依次加入涉及生理性肝发生的诱导因子诱导其沿着肝谱系分化。分化过程基本一致，细胞培养物逐渐表达越来越多的肝谱系标志物，包括GATA 4、HNF 4 α、甲胎蛋白、CD 26、白蛋白、α-1-抗胰蛋白酶、Cyp 7A 1和Cyp 3A 4。肝细胞显示功能性肝特征，如糖原储存、吲哚菁绿色摄取和释放以及白蛋白分泌。在急性肝损伤的小鼠模型中，hESC衍生的定形内胚层分化成肝细胞并重新填充受损的肝脏。这里描述的方法代表了有效产生肝细胞用于再生医学和药物发现的重要一步。
Differentiation of human embryonic stem cells (hESCs) to specific functional cell types can be achieved using methods that mimic in vivo embryonic developmental programs. Current protocols for generating hepatocytes from hESCs are hampered by inefficient differentiation procedures that lead to low yields and large cellular heterogeneity. We report here a robust and highly efficient process for the generation of high-purity (70%) hepatocyte cultures from hESCs that parallels sequential hepatic development in vivo. Highly enriched populations of definitive endoderm were generated from hESCs and then induced to differentiate along the hepatic lineage by the sequential addition of inducing factors implicated in physiological hepatogenesis. The differentiation process was largely uniform, with cell cultures progressively expressing increasing numbers of hepatic lineage markers, including GATA4, HNF4 alpha, alpha-fetoprotein, CD26, albumin, alpha-1-antitrypsin, Cyp7A1, and Cyp3A4. The hepatocytes exhibited functional hepatic characteristics, such as glycogen storage, indocyanine green uptake and release, and albumin secretion. In a mouse model of acute liver injury, the hESC-derived definitive endoderm differentiated into hepatocytes and repopulated the damaged liver. The methodology described here represents a significant step toward the efficient generation of hepatocytes for use in regenerative medicine and drug discovery.