Laboratory of Molecular Cell Biology, Institute of Biochemistry and Cell Biology, Shanghai Institutes for Biological Sciences, Shanghai, China.
Gastroenterology. 2010 Dec;139(6):2158-2169.e8. doi: 10.1053/j.gastro.2010.08.042. Epub 2010 Aug 27.
BACKGROUND & AIMS: Hepatocyte-like cells can be derived from pluripotent stem cells such as embryonic stem (ES) cells, but ES cell-derived hepatic cells with extensive capacity to repopulate liver have not been identified. We aimed to identify and purify ES cell-derived hepatoblast-like progenitor cells and to explore their capacity for liver repopulation in mice after in vitro expansion.
Unmanipulated mouse ES cells were cultured under defined conditions and allowed to undergo stepwise hepatic differentiation. The derived hepatic cells were examined by morphologic, fluorescence-activated cell sorting, gene expression, and clonal expansion analyses. The capacities of ES cell-derived hepatic progenitor cells to repopulate liver were investigated in mice that were deficient in fumarylacetoacetate hydrolase (Fah) (a model of liver injury).
Mouse ES cells were induced to differentiate into a population that contained hepatic progenitor cells; this population included cells that expressed epithelial cell adhesion molecule (EpCAM) but did not express c-Kit. Clonal hepatic progenitors that arose from single c-Kit(-)EpCAM(+) cells could undergo long-term expansion and maintain hepatoblast-like characteristics. Enriched c-Kit(-)EpCAM(+) cells and clonally expanded hepatic progenitor cells repopulated the livers of Fah-deficient mice without inducing tumorigenesis.
ES cell-derived c-Kit(-)EpCAM(+) cells contain a population of hepatoblast-like progenitor cells that can repopulate livers of mice.
肝样细胞可由多能干细胞如胚胎干细胞（ES 细胞）衍生而来，但尚未鉴定出具有广泛肝重编程能力的 ES 细胞衍生的肝细胞。我们旨在鉴定和纯化 ES 细胞衍生的肝前体细胞样祖细胞，并探讨其在体外扩增后在小鼠体内肝重编程的能力。
未处理的小鼠 ES 细胞在特定条件下培养，并允许其进行逐步肝分化。通过形态学、荧光激活细胞分选、基因表达和克隆扩增分析来检查衍生的肝细胞。在缺乏延胡索酰乙酰乙酸水解酶（Fah）的小鼠（肝损伤模型）中研究 ES 细胞衍生的肝祖细胞肝重编程的能力。
小鼠 ES 细胞被诱导分化为包含肝祖细胞的群体；该群体包括表达上皮细胞黏附分子（EpCAM）但不表达 c-Kit 的细胞。源自单个 c-Kit(-)EpCAM(+)细胞的克隆肝祖细胞可以进行长期扩增并保持肝前体细胞样特征。富含 c-Kit(-)EpCAM(+)细胞和克隆扩增的肝祖细胞可重编程 Fah 缺陷小鼠的肝脏，而不会诱导肿瘤发生。
ES 细胞衍生的 c-Kit(-)EpCAM(+)细胞包含一群具有肝重编程能力的肝前体细胞样祖细胞。
