The object is to obtain a highly functional hepatocyte that has a high ammonia-metabolizing function and is theoretically able to infinitely proliferate from a human iPS cell. There is provided a method for preparing an artificial hepatocyte comprising the following steps: the differentiation induction step 1 of culturing an iPS cell in a differentiation induction medium I containing activin A; the differentiation induction step 2 of culturing a cell obtained in the differentiation induction step 1 in a differentiation induction medium II containing bone morphogenetic protein 4 (BMP4) and fibroblast growth factor 2 (FGF2); and the differentiation induction step 3 of culturing a cell obtained in the differentiation induction step 2 in a differentiation induction medium III containing hepatocyte growth factor (HGF), oncostatin M, dexamethasone, and N,N'-(methylenebis)(4,1-phenylene)diacetamide (FH1) and/or 2-([N-(5-chloro-2-methylphenyl)(methylsulfonamido)-N-(2,6-difluorophenyl)acetamide (FPH1) to obtain an artificial hepatocyte. The obtained artificial hepatocyte has an ammonia-metabolizing function of 100 µg/dl/24h or higher, and can constitute a mesh structure.
目的是获得一种高功能肝细胞,该肝细胞具有高
氨代谢功能,理论上能够从人类 iPS 细胞无限增殖。本发明提供了一种制备人工肝细胞的方法,包括以下步骤:分化诱导步骤 1:在含有活化素 A 的分化诱导
培养基 I 中培养 iPS 细胞; 分化诱导步骤 2:在含有骨形态发生蛋白 4(
BMP4)和成纤维细胞生长因子 2(FGF2)的分化诱导
培养基 II 中培养在分化诱导步骤 1 中获得的细胞;分化诱导步骤 3:在分化诱导
培养基 III 中培养分化诱导步骤 2 中获得的细胞,该
培养基 III 含有
肝细胞生长因子(
HGF)、昂科司他丁 M、N,N'-(亚甲基双)(4,1-亚苯基)二乙酰胺(FH1)和/或 2-([N-(5-
氯-2-甲基苯基)(甲磺酰胺基)-N-(2,6-二
氟苯基)乙酰胺(FPH1)),以获得人造肝细胞。获得的人工肝细胞的
氨代谢功能为 100 µg/dl/24h 或更高,并可构成网状结构。