Generation of Human Islets Through Expansion and Differentiation of Non-islet Pancreatic Cells Discarded (Pancreatic Discard) After Islet Isolation

• DOI: 10.1097/01.mpa.0000202945.78331.93
• Publication Date: 2006-03-21T09:13:21Z
• Authors: Ivan Todorov; Keiko Omori; Michael Pascual; Jeffery Rawson; Indu Nair; Luis Valiente; Tommy Vuong; Takeru Matsuda; Chris Orr; Kevin Ferreri; Craig V. Smith; Fouad Kandeel; Yoko Mullen
• Keywords: Transdifferentiation

Objectives: Islet transplantation is hampered by the shortage of donor tissues. Our objective was to generate islet-like cell clusters (ICCs) from cultures non-islet pancreatic cells. Methods: The starting cultured cells came fractions human pancreases after enzymatic digestion and purification for purpose islet isolation. Initially, these expanded in monolayer became confluent on collagen-coated flasks. After trypsination suspension a defined differentiation medium, aggregated form ICCs. Results: initial population consisted less than 1% insulin-positive cells, 44% amylase-positive 41% cytokeratin (CK) 7-positive, or CK19+ but PDX-1+ were absent. Cells later stages showed signs dedifferentiation/transdifferentiation. At time harvesting, more 90% positive CK 7/19 PDX-1, insulin-positive. aggregation, ICCs appeared redifferentiated, contained glucose-responsive, insulin-secreting with an insulin content measuring 20% that found freshly isolated islets same pancreas. transplanted into athymic mice removed 4 months did acquire morphology mature islets, indicating further maturation vivo transplantation. Human C-peptide detected recipient animal sera. Conclusion: Using specified culture methods, pancreas can resembling islets. These ICCs, obtained are otherwise discarded, continue differentiate become