In vitro hepatic maturation of human embryonic stem cells by using a mesenchymal cell line derived from murine fetal livers

Takamichi Ishii, Kentaro Yasuchika, Ken Fukumitsu, Tatsuya Kawamoto, Miho Kawamura-Saitoh, Yuji Amagai, Iwao Ikai, Shinji Uemoto, Eihachiro Kawase, Hirofumi Suemori, Norio Nakatsuji
Cell and Tissue Research 2010, 339 (3): 505-12
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.

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