title: Enrichment of Pluripotent Stem Cell-Derived Hepatocyte-Like Cells by Ammonia Treatment.
abstract: Embryonic stem cells (ESCs) and induced pluripotent stem cells (iPSCs) are potential resources for the regeneration of defective organs, including the liver. However, some obstacles must be overcome before this becomes reality. Undifferentiated cells that remain following differentiation have teratoma-forming potential. Additionally, practical applications require a large quantity of differentiated cells, so the differentiation process must be economical. Here we describe a DNA microarray-based global analysis of the gene expression profiles of differentiating human pluripotent stem cells. We identified differences and commonalities among six human pluripotent stem cell lines: the hESCs KhES1, KhES2, KhES3, and H1, and the iPSCs 201B7 and 243G1. Embryoid bodies (EBs) formed without requiring supplementation with inducing factors. EBs also expressed some liver-specific metabolic genes including the ammonia-metabolizing enzymes glutamine synthetase and carbamoyl-phosphate synthase 1. Real-time PCR analysis revealed hepatocyte-like differentiation of EBs treated with ammonia in Lanford medium. Analysis of DNA microarray data suggested that hepatocyte-like cells were the most abundant population in ammonia-treated cells. Furthermore, expression levels of undifferentiated pluripotent stem cell markers were drastically reduced, suggesting a reduced teratoma-forming capacity. These results indicate that treatment of EBs with ammonia in Lanford medium may be an effective inducer of hepatic differentiation in absence of expensive inducing factors.
authors.affiliation: Department of Biotechnology and Biomedical Engineering, Institute for Biomedical Sciences, Interdisciplinary Cluster for Cutting Edge Research, Shinshu University Matsumoto, 3-1-1 Asahi, Matsumoto 390-8621, Japan.
authors.forename: Daihachiro
authors.initials: D
authors.lastname: Tomotsune
authors.affiliation: Department of Histology and Embryology, Shinshu University School of Medicine, 3-1-1 Asahi, Matsumoto 390-8621, Japan.
authors.forename: Kanji
authors.initials: K
authors.lastname: Hirashima
authors.affiliation: Department of Histology and Embryology, Shinshu University School of Medicine, 3-1-1 Asahi, Matsumoto 390-8621, Japan.
authors.forename: Masako
authors.initials: M
authors.lastname: Fujii
authors.affiliation: Department of Biotechnology and Biomedical Engineering, Institute for Biomedical Sciences, Interdisciplinary Cluster for Cutting Edge Research, Shinshu University Matsumoto, 3-1-1 Asahi, Matsumoto 390-8621, Japan.|Department of Histology and Embryology, Shinshu University School of Medicine, 3-1-1 Asahi, Matsumoto 390-8621, Japan.
authors.forename: Fengming
authors.initials: F
authors.lastname: Yue
authors.affiliation: Nissui Pharmaceutical Co., Ltd., 1075-2 Hokunanmoro, Yuki, Ibaraki 307-0036, Japan.
authors.forename: Ken
authors.initials: K
authors.lastname: Matsumoto
authors.affiliation: Bourbon Corporation, 4-2-14 Matsunami, Kashiwazaki, Niigata 945-8611, Japan.
authors.forename: Sakiko
authors.initials: S
authors.lastname: Takizawa-Shirasawa
authors.affiliation: Bourbon Corporation, 4-2-14 Matsunami, Kashiwazaki, Niigata 945-8611, Japan.
authors.forename: Tadayuki
authors.initials: T
authors.lastname: Yokoyama
authors.affiliation: Department of Biotechnology and Biomedical Engineering, Institute for Biomedical Sciences, Interdisciplinary Cluster for Cutting Edge Research, Shinshu University Matsumoto, 3-1-1 Asahi, Matsumoto 390-8621, Japan.|Department of Histology and Embryology, Shinshu University School of Medicine, 3-1-1 Asahi, Matsumoto 390-8621, Japan.
authors.forename: Katsunori
authors.initials: K
authors.lastname: Sasaki
mesh_terms: D000641:Ammonia / Q000494:pharmacology*; D000818:Animals; D002454:Cell Differentiation; D019070:Cell Lineage; D022781:Hepatocytes / Q000166:cytology / Q000187:drug effects*; D006801:Humans; D020411:Oligonucleotide Array Sequence Analysis; D039904:Pluripotent Stem Cells / Q000166:cytology / Q000187:drug effects*
chemical_list: D000641:Ammonia