title: High oxygen condition facilitates the differentiation of mouse and human pluripotent stem cells into pancreatic progenitors and insulin-producing cells.
abstract: Pluripotent stem cells have potential applications in regenerative medicine for diabetes. Differentiation of stem cells into insulin-producing cells has been achieved using various protocols. However, both the efficiency of the method and potency of differentiated cells are insufficient. Oxygen tension, the partial pressure of oxygen, has been shown to regulate the embryonic development of several organs, including pancreatic β-cells. In this study, we tried to establish an effective method for the differentiation of induced pluripotent stem cells (iPSCs) into insulin-producing cells by culturing under high oxygen (O2) conditions. Treatment with a high O2 condition in the early stage of differentiation increased insulin-positive cells at the terminus of differentiation. We found that a high O2 condition repressed Notch-dependent gene Hes1 expression and increased Ngn3 expression at the stage of pancreatic progenitors. This effect was caused by inhibition of hypoxia-inducible factor-1α protein level. Moreover, a high O2 condition activated Wnt signaling. Optimal stage-specific treatment with a high O2 condition resulted in a significant increase in insulin production in both mouse embryonic stem cells and human iPSCs and yielded populations containing up to 10% C-peptide-positive cells in human iPSCs. These results suggest that culturing in a high O2 condition at a specific stage is useful for the efficient generation of insulin-producing cells.
authors.affiliation: From the Department of Molecular Physiology, Faculty of Life Sciences, Kumamoto University, 1-1-1 Honjo, Chuo-ku, Kumamoto 860-8556, Japan.
authors.forename: Farzana
authors.initials: F
authors.lastname: Hakim
authors.forename: Taku
authors.initials: T
authors.lastname: Kaitsuka
authors.forename: Jamiruddin Mohd
authors.initials: JM
authors.lastname: Raeed
authors.forename: Fan-Yan
authors.initials: FY
authors.lastname: Wei
authors.forename: Nobuaki
authors.initials: N
authors.lastname: Shiraki
authors.forename: Tadayuki
authors.initials: T
authors.lastname: Akagi
authors.forename: Takashi
authors.initials: T
authors.lastname: Yokota
authors.forename: Shoen
authors.initials: S
authors.lastname: Kume
authors.forename: Kazuhito
authors.initials: K
authors.lastname: Tomizawa
mesh_terms: D000818:Animals; D002454:Cell Differentiation / Q000187:drug effects* / Q000502:physiology; D002460:Cell Line; D053595:Embryonic Stem Cells / Q000166:cytology / Q000378:metabolism*; D006801:Humans; D051795:Hypoxia-Inducible Factor 1, alpha Subunit / Q000378:metabolism; D007328:Insulin / Q000096:biosynthesis*; D050417:Insulin-Secreting Cells / Q000166:cytology / Q000378:metabolism*; D051379:Mice; D010100:Oxygen / Q000378:metabolism / Q000494:pharmacology*; D039904:Pluripotent Stem Cells / Q000166:cytology / Q000378:metabolism*
chemical_list: C497442:HIF1A protein, human; C497443:Hif1a protein, mouse; D051795:Hypoxia-Inducible Factor 1, alpha Subunit; D007328:Insulin; D010100:Oxygen
keywords: Differentiation; Embryonic Stem Cell; Hypoxia-inducible Factor; Induced Pluripotent Stem Cells; Regenerative Medicine