title: Enhanced oxygenation promotes beta-cell differentiation in vitro.
abstract: Despite progress in our knowledge about pancreatic islet specification, most attempts at differentiating stem/progenitor cells into functional, transplantable beta cells have met only with moderate success thus far. A major challenge is the intrinsic simplicity of in vitro culture systems, which cannot approximate the physiological complexity of in vivo microenvironments. Oxygenation is a critical limitation of standard culture methods, and one of special relevance for the development of beta cells, known for their high O(2) requirements. Based on our understanding of islet physiology, we have tested the hypothesis that enhanced O(2) delivery (as provided by novel perfluorocarbon-based culture devices) may result in higher levels of beta-cell differentiation from progenitor cells in vitro. Using a mouse model of pancreatic development, we demonstrate that a physiological-like mode of O(2) delivery results in a very significant upregulation of endocrine differentiation markers (up to 30-fold for insulin one and 2), comparable to relevant in vivo controls. This effect was not observed by merely increasing environmental O(2) concentrations in conventional settings. Our findings indicate that O(2) plays an important role in the differentiation of beta cells from their progenitors and may open the door to more efficient islet differentiation protocols from embryonic and/or adult stem cells. Disclosure of potential conflicts of interest is found at the end of this article.
authors.affiliation: Diabetes Research Institute, University of Miami Leonard M. Miller School of Medicine, Miami, Florida 33136, USA.
authors.forename: Christopher A
authors.initials: CA
authors.lastname: Fraker
authors.forename: Silvia
authors.initials: S
authors.lastname: Alvarez
authors.forename: Panagiotis
authors.initials: P
authors.lastname: Papadopoulos
authors.forename: Jaime
authors.initials: J
authors.lastname: Giraldo
authors.forename: Weiyong
authors.initials: W
authors.lastname: Gu
authors.forename: Camillo
authors.initials: C
authors.lastname: Ricordi
authors.forename: Luca
authors.initials: L
authors.lastname: Inverardi
authors.forename: Juan
authors.initials: J
authors.lastname: Domínguez-Bendala
mesh_terms: D000818:Animals; D002454:Cell Differentiation / Q000502:physiology*; D002478:Cells, Cultured; D005260:Female; D050417:Insulin-Secreting Cells / Q000166:cytology* / Q000378:metabolism*; D051379:Mice; D008810:Mice, Inbred C57BL; D008808:Mice, Inbred CBA; D010100:Oxygen / Q000378:metabolism* / Q000502:physiology; D010101:Oxygen Consumption / Q000502:physiology*; D011247:Pregnancy
chemical_list: D010100:Oxygen