Method for isolation of mouse pancreatic stem cells

Replacement of beta-cell mass offers an alternative to standard insulin treatment for diabetes and may overcome the long-term side effects associated with current therapies. Pancreatic stem/progenitor cells could become a useful target for beta-cell replacement therapy in diabetic patients. We have established a method for isolating mouse pancreatic stem cells. In this study, pancreatic stem cells were isolated from 8-week-old mice. After purification on a density gradient, the density range of 1.062-1.11 contained pancreatic stem cells. The islets from the layers were deleted by dithizone staining and hand-picking under a dissecting microscope. The remnant cells were then cultured, inoculated into 96-well plates, and cloned by limiting dilution. One of the wells contained cells, named HN#5 cells, which expressed ductal cell markers, such as cytokeratin-19. HN#5 cells differentiated into insulin-producing cells and albumin-producing cells by induction medium. The isolation technique described here may be useful for identification and isolation of human pancreatic stem/progenitor cells.