<i>In vitro</i> cultivation of human islets from expanded ductal tissue

• Susan Bonner-Weir

  Joslin Diabetes Center, Department of Medicine, Harvard Medical School, Boston, MA 02215

• Monica Taneja

  Joslin Diabetes Center, Department of Medicine, Harvard Medical School, Boston, MA 02215

• Gordon C. Weir

  Joslin Diabetes Center, Department of Medicine, Harvard Medical School, Boston, MA 02215

• Krystyna Tatarkiewicz

  Joslin Diabetes Center, Department of Medicine, Harvard Medical School, Boston, MA 02215

• Ki-Ho Song

  Joslin Diabetes Center, Department of Medicine, Harvard Medical School, Boston, MA 02215

• Arun Sharma

  Joslin Diabetes Center, Department of Medicine, Harvard Medical School, Boston, MA 02215

• John J. O'Neil

  Joslin Diabetes Center, Department of Medicine, Harvard Medical School, Boston, MA 02215

抄録

A major obstacle to successful islet transplantation for both type 1 and 2 diabetes is an inadequate supply of insulin-producing tissue. This need for transplantable human islets has stimulated efforts to expand existing pancreatic islets and/or grow new ones. To test the hypothesis that human adult duct tissue could be expanded and differentiated in vitro to form islet cells, digested pancreatic tissue that is normally discarded from eight human islet isolations was cultured under conditions that allowed expansion of the ductal cells as a monolayer whereupon the cells were overlaid with a thin layer of Matrigel. With this manipulation, the monolayer of epithelial cells formed three-dimensional structures of ductal cysts from which 50-to 150-μm diameter islet-like clusters of pancreatic endocrine cells budded. Over 3–4 weeks culture the insulin content per flask increased 10- to 15-fold as the DNA content increased up to 7-fold. The cultivated human islet buds were shown by immunofluorescence to consist of cytokeratin 19-positive duct cells and hormone-positive islet cells. Double staining of insulin and non-β cell hormones in occasional cells indicated immature cells still in the process of differentiation. Insulin secretion studies were done over 24 h in culture. Compared with their basal secretion at 5 mM glucose, cysts/cultivated human islet buds exposed to stimulatory 20 mM glucose had a 2.3-fold increase in secreted insulin. Thus, duct tissue from human pancreas can be expanded in culture and then be directed to differentiate into glucose responsive islet tissue in vitro . This approach may provide a potential new source of pancreatic islet cells for transplantation.

収録刊行物

• Proceedings of the National Academy of Sciences

  Proceedings of the National Academy of Sciences 97 (14), 7999-8004, 2000-07-05

  Proceedings of the National Academy of Sciences

キーワード

• Multidisciplinary

詳細情報

• CRID: 1363951795574038656
• NII論文ID: 80011918681
• DOI: 10.1073/pnas.97.14.7999
• ISSN: 10916490
• ISSN: 00278424
• Web Site: https://pnas.org/doi/pdf/10.1073/pnas.97.14.7999
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