Abstract
BackgroundIn 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 FindingsIn 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 multiplicity of infection, we created a chimeric graft with a subpopulation of human beta cells expressing GFP and found both GFP-positive and GFP-negative beta cells within single islets.ConclusionThe detection of both labeled and unlabeled beta cells in single islets demonstrates that beta cells present in a human islet are derived from multiple progenitors thus providing the first dynamic analysis of human islet formation during development. This human transgenic-like tool can be widely used to elucidate dynamic genetic processes in human tissue formation.
Highlights
The mature pancreas consists of acinar cells that produce digestive enzymes secreted via the ducts into the intestine and hormone-producing endocrine cells
We have previously shown that islet formation occurs when human fetal pancreases are grafted under the kidney capsule of scid mice [16,20]
We first compared the cyto-architecture of islets that developed from human fetal pancreas transplanted in scid mice to that of islets found in human and mouse pancreas
Summary
The mature pancreas consists of acinar cells that produce digestive enzymes secreted via the ducts into the intestine and hormone-producing endocrine cells. The endocrine cells are organized into micro-organs called islets of Langerhans. Rodent islet cell development is well studied [1,2], little is known about islet formation in humans. 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. Little is known about human islet formation. 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
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