In vitro hepatic maturation of human embryonic stem cells by using a mesenchymal cell line derived from murine fetal livers
In vitro hepatic maturation of human embryonic stem cells by using a mesenchymal cell line derived from murine fetal livers
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DOI：
10.1007/s00441-009-0906-7
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发表时间：
2010-03-01
影响因子：
3.6
通讯作者：
Nakatsuji, Norio
中科院分区：
文献类型：
--
作者：
Ishii, Takamichi;Yasuchika, Kentaro;Nakatsuji, Norio
人胚胎干细胞（hESCs）衍生的肝细胞是一种有吸引力的再生医学细胞来源。我们之前报道了利用细胞外基质和生长因子将hESCs分化为产生甲胎蛋白（AFP）的内胚层细胞。我们还报道了MLSgt20细胞系的建立，该细胞系来源于小鼠胎儿肝脏中的间充质细胞，并加速了小鼠肝祖细胞和小鼠ESCs的肝脏成熟。在本研究中，利用流式细胞仪分离hesc来源的afp产生细胞，并与MLSgt20细胞共培养。共培养的hesc来源的产afp细胞具有肝细胞的免疫细胞学特征，表达成熟的肝细胞标志物（通过逆转录和聚合酶链反应表明），并表现出更高的肝细胞功能，包括氨去除，细胞色素P450 3A4/7活性，以及产生和储存糖原的能力。然而，MLSgt20细胞不会直接导致未分化的hESCs成熟为肝细胞样细胞。因此，共培养方法成功地诱导hesc来源的内胚层细胞向功能性肝细胞样细胞分化。
Hepatocytes derived from human embryonic stem cells (hESCs) are an attractive cell source for regenerative medicine. We previously reported the differentiation of hESCs into alpha-fetoprotein (AFP)-producing endodermal cells by using extracellular matrix and growth factors. We also reported the establishment of the MLSgt20 cell line, which was derived from mesenchymal cells residing in murine fetal livers and accelerated the hepatic maturation of both murine hepatic progenitor cells and murine ESCs. In this study, hESC-derived AFP-producing cells were isolated by using a flow cytometer and co-cultured with MLSgt20 cells. The co-cultured hESC-derived AFP-producing cells had the immunocytological characteristics of hepatocytes, expressed mature hepatocyte markers (as indicated by reverse transcription and the polymerase chain reaction), and displayed higher hepatocyte functions including ammonia removal, cytochrome P450 3A4/7 activity, and the ability to produce and store glycogen. However, the MLSgt20 cells did not directly cause undifferentiated hESCs to mature into hepatocyte-like cells. The co-culture method was thus successfully shown to induce the differentiation of hESC-derived endodermal cells into functional hepatocyte-like cells.