beta cells within single human islets originate from multiple progenitors在单一人类胰岛β细胞来源于多个祖细胞.pdfVIP

Beta Cells within Single Human Islets Originate from Multiple Progenitors ¨ 1 2 2 3 3 Raphael Scharfmann , Xiangwei Xiao , Harry Heimberg , Jacques Mallet , Philippe Ravassard * 1 University Paris-Descartes, Faculty of Medicine, INSERM, Necker Hospital, U845, Paris, France, 2 Diabetes Research Center, Vrije Universiteit Brussel, Brussels, Belgium, ´ 3 Centre National de la Recherche Scientifique and Universite Pierre et Marie Curie, LGN UMR 7091, Paris, France Abstract Background: In both humans and rodents, glucose homeostasis is controlled by micro-organs called islets of Langerhans composed of beta cells, associated with other endocrine cell types. Most of our understanding of islet cell differentiation and morphogenesis is derived from rodent developmental studies. However, little is known about human islet formation. The lack of adequate experimental models has restricted the study of human pancreatic development to the histological analysis of different stages of pancreatic development. Our objective was to develop a new experimental model to (i) transfer genes into developing human pancreatic cells and (ii) validate gene transfer by defining the clonality of developing human islets. Methods and Findings: In this study, a unique model was developed combining ex vivo organogenesis from human fetal pancreatic tissue and cell type-specific lentivirus-mediated gene transfer. Human pancreatic progenitors were transduced with lentiviruses expressing GFP under the control of an insulin promoter and grafted to severe combined immunodeficient mice, allowing human beta cell differentiation and islet morphogenesis. By performing gene transfer at low multiplici