title: Erythropoietin facilitates definitive endodermal differentiation of mouse embryonic stem cells via activation of ERK signaling.
abstract: Artificially generated pancreatic β-cells from pluripotent stem cells are expected for cell replacement therapy for type 1 diabetes. Several strategies are adopted to direct pluripotent stem cells toward pancreatic differentiation. However, a standard differentiation method for clinical application has not been established. It is important to develop more effective and safer methods for generating pancreatic β-cells without toxic or mutagenic chemicals. In the present study, we screened several endogenous factors involved in organ development to identify the factor, which induced the efficiency of pancreatic differentiation and found that treatment with erythropoietin (EPO) facilitated the differentiation of mouse embryonic stem cells (ESCs) into definitive endoderm. At an early stage of differentiation, EPO treatment significantly increasedSox17gene expression, as a marker of the definitive endoderm. Contrary to the canonical function of EPO, it did not affect the levels of phosphorylated JAK2 and STAT5, but stimulated the phosphorylation of ERK1/2 and Akt. The MEK inhibitor U0126 significantly inhibited EPO-inducedSox17expression. The differentiation of ESCs into definitive endoderm is an important step for the differentiation into pancreatic and other endodermal lineages. This study suggests a possible role of EPO in embryonic endodermal development and a new agent for directing the differentiation into endodermal lineages like pancreatic β-cells.
authors.affiliation: Department of Molecular Physiology, Faculty of Life Sciences, Kumamoto University, Kumamoto, Japan.
authors.forename: Taku
authors.initials: T
authors.lastname: Kaitsuka
authors.affiliation: Department of Molecular Physiology, Faculty of Life Sciences, Kumamoto University, Kumamoto, Japan.
authors.forename: Kohei
authors.initials: K
authors.lastname: Kobayashi
authors.affiliation: Department of Molecular Physiology, Faculty of Life Sciences, Kumamoto University, Kumamoto, Japan.
authors.forename: Wakako
authors.initials: W
authors.lastname: Otsuka
authors.affiliation: Department of Molecular Physiology, Faculty of Life Sciences, Kumamoto University, Kumamoto, Japan.
authors.forename: Takuya
authors.initials: T
authors.lastname: Kubo
authors.affiliation: Department of Molecular Physiology, Faculty of Life Sciences, Kumamoto University, Kumamoto, Japan.
authors.forename: Farzana
authors.initials: F
authors.lastname: Hakim
authors.affiliation: Department of Molecular Physiology, Faculty of Life Sciences, Kumamoto University, Kumamoto, Japan.
authors.forename: Fan-Yan
authors.initials: FY
authors.lastname: Wei
authors.affiliation: Department of Stem Cell Biology, Institute of Molecular Embryology and Genetics, Kumamoto University, Kumamoto, Japan; and.|Graduate School of Bioscience and Biotechnology, Tokyo Institute of Technology, Yokohama, Japan.
authors.forename: Nobuaki
authors.initials: N
authors.lastname: Shiraki
authors.affiliation: Department of Stem Cell Biology, Institute of Molecular Embryology and Genetics, Kumamoto University, Kumamoto, Japan; and.|Graduate School of Bioscience and Biotechnology, Tokyo Institute of Technology, Yokohama, Japan.
authors.forename: Shoen
authors.initials: S
authors.lastname: Kume
authors.affiliation: Department of Molecular Physiology, Faculty of Life Sciences, Kumamoto University, Kumamoto, Japan; tomikt@kumamoto-u.ac.jp.
authors.forename: Kazuhito
authors.initials: K
authors.lastname: Tomizawa
mesh_terms: D000818:Animals; D002454:Cell Differentiation / Q000502:physiology; D002460:Cell Line; D053595:Embryonic Stem Cells / Q000166:cytology* / Q000502:physiology*; D004707:Endoderm / Q000166:cytology* / Q000502:physiology; D004921:Erythropoietin / Q000378:metabolism*; D018507:Gene Expression Regulation, Developmental / Q000502:physiology; D050417:Insulin-Secreting Cells / Q000166:cytology* / Q000502:physiology; D020935:MAP Kinase Signaling System / Q000502:physiology*; D051379:Mice
chemical_list: D004921:Erythropoietin
keywords: ERK; definitive endoderm; differentiation; embryonic stem cell; erythropoietin