Enhanced oxygenation promotes beta cell differentiation in vitro

¹ Diabetes Research Institute, University of Miami Leonard M. Miller School of Medicine, 1450 NW 10^th Avenue, Miami, FL 33136; Department of Biomedical Engineering; University of Miami, P.O. Box 248294, Coral Gables, FL 33124
² Diabetes Research Institute, University of Miami Leonard M. Miller School of Medicine, 1450 NW 10^th Avenue, Miami, FL 33136
³ Department of Biomedical Engineering; University of Miami, P.O. Box 248294, Coral Gables, FL 33124

^* To whom correspondence should be addressed. E-mail: jdominguez2{at}med.miami.edu.

	Abstract

Despite progress in our knowledge about pancreatic islet specification, most attempts at differentiating stem/progenitor cells into functional, transplantable 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 cells, known for their high O₂ requirements. Based on our understanding of islet physiology, we have tested the hypothesis that enhanced O₂ delivery (as provided by novel perfluorocarbon-based culture devices) may result in higher levels of cell differentiation from progenitor cells in vitro. Using a mouse model of pancreatic development, we demonstrate that a physiological-like mode of O₂ delivery results in a very significant up-regulation of endocrine differentiation markers (up to 30-fold for insulin 1 and 2), comparable to relevant in vivo controls. This effect was not observed by merely increasing environmental O₂ concentrations in conventional settings. Our findings indicate that O₂ plays an important role in the differentiation of cells from their progenitors, and may open the door to more efficient islet differentiation protocols from embryonic and/or adult stem cells.