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Abstract
Ammonia has a cytotoxic effect and can therefore be used as a selection agent for enrichment of zone I hepatocytes. However, it has not yet been determined whether ammonia-treated hepatocyte-like cells are able to proliferate in vitro. In this study, we employed an ammonia selection strategy to purify hepatocyte-like cells that were differentiated from human embryonic stem cells (ESCs) and from induced pluripotent stem cells (iPSCs). The resistance to cytotoxicity or cell death by ammonia is likely attributable to the metabolism of ammonia in the cells. In addition to ammonia metabolism-related genes, ammonia-selected hepatocytes showed increased expression of the cytochrome P450 genes. Additionally, the ammonia-selected cells achieved immortality or at least an equivalent life span to human pluripotent stem cells without affecting expression of the liver-associated genes. Ammonia treatment in combination with in vitro propagation is useful for obtaining large quantities of hepatocytes.
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1 National Center for Child Health and Development Research Institute, Center for Regenerative Medicine, Tokyo, Japan (GRID:grid.63906.3a) (ISNI:0000 0004 0377 2305); Tokyo University of Pharmacy and Life Sciences, Laboratory of Molecular Neuroscience and Neurology, Hachioji, Tokyo, Japan (GRID:grid.410785.f) (ISNI:0000 0001 0659 6325)
2 National Center for Child Health and Development Research Institute, Center for Regenerative Medicine, Tokyo, Japan (GRID:grid.63906.3a) (ISNI:0000 0004 0377 2305); Tokyo Medical University, Department of Molecular Pathology, Tokyo, Japan (GRID:grid.410793.8) (ISNI:0000 0001 0663 3325)
3 National Center for Child Health and Development Research Institute, Center for Regenerative Medicine, Tokyo, Japan (GRID:grid.63906.3a) (ISNI:0000 0004 0377 2305)
4 National Center for Child Health and Development Research Institute, Center for Regenerative Medicine, Tokyo, Japan (GRID:grid.63906.3a) (ISNI:0000 0004 0377 2305); Tokyo Metropolitan Institute of Gerontology, Research Team for Geriatric Medicine (Vascular Medicine), Tokyo, Japan (GRID:grid.420122.7) (ISNI:0000 0000 9337 2516)
5 Kitasato University School of Science, Laboratory of Stem Cell Biology, Department of Biosciences, Kanagawa, Japan (GRID:grid.410786.c) (ISNI:0000 0000 9206 2938)
6 Tokyo Medical University, Department of Molecular Pathology, Tokyo, Japan (GRID:grid.410793.8) (ISNI:0000 0001 0663 3325)
7 National Center for Child Health and Development Research Institute, Center for Regenerative Medicine, Tokyo, Japan (GRID:grid.63906.3a) (ISNI:0000 0004 0377 2305); Saitama Children’s Medical Center, Saitama, Japan (GRID:grid.416697.b) (ISNI:0000 0004 0569 8102)
8 National Center for Child Health and Development, Organ Transplantation Center, Tokyo, Japan (GRID:grid.63906.3a) (ISNI:0000 0004 0377 2305)
9 Tokai University School of Medicine, Department of Molecular Life Sciences, Isehara, Japan (GRID:grid.265061.6) (ISNI:0000 0001 1516 6626)
10 National Cancer Center, Project for Prevention of HPV-Related Cancer, Exploratory Oncology Research and Clinical Trial Center, Chiba, Japan (GRID:grid.272242.3) (ISNI:0000 0001 2168 5385)
11 Tokyo University of Pharmacy and Life Sciences, Laboratory of Molecular Neuroscience and Neurology, Hachioji, Tokyo, Japan (GRID:grid.410785.f) (ISNI:0000 0001 0659 6325)