1. Ammonia-based enrichment and long-term propagation of zone I hepatocyte-like cells
- Author
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Akihiro Umezawa, Hidenori Nonaka, Tohru Kiyono, Saeko Akiyama, Masahiko Kuroda, Shoko Miyata, Akihide Kamiya, Palaksha Kanive Javaregowda, Noriaki Saku, Junji Yamauchi, Kenta Ite, Atsuko Nakazawa, Tohru Kimura, Mureo Kasahara, Nagisa Takashima, Masashi Toyoda, and Ruri Tsuneishi
- Subjects
Cell biology ,Programmed cell death ,Science ,Induced Pluripotent Stem Cells ,Stem cells ,Article ,Mice ,Ammonia ,medicine ,Animals ,Humans ,Cytotoxic T cell ,Cytotoxicity ,Induced pluripotent stem cell ,Cells, Cultured ,Embryonic Stem Cells ,Cell Proliferation ,Multidisciplinary ,Chemistry ,Embryonic stem cell ,In vitro ,medicine.anatomical_structure ,Hepatocyte ,Hepatocytes ,Medicine ,Stem cell - 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.
- Published
- 2021