Differentiation of Sendai Virus-Reprogrammed iPSC into beta Cells, Compared with Human Pancreatic Islets and Immortalized beta Cell Line

摘要

Background: New sources of insulin-secreting cells are strongly in demand for treatment of diabetes. Induced pluripotent stem cells (iPSCs) have the potential to generate insulin-producing cells (i beta). However, the gene expression profile and secretory function of i beta still need to be validated in comparison with native beta cells. Methods: Two clones of human iPSCs, reprogrammed from adult fibroblasts through integration-free Sendai virus, were differentiated into i and compared with donor pancreatic islets and EndoC-beta H1, an immortalized human beta cell line. Results: Both clones of iPSCs differentiated into insulin(+) cells with high efficiency (up to 20%). i beta were negative for pluripotency markers (Oct4, Sox2, Ssea4) and positive for Pdx1, Nkx6.1, Chromogranin A, PC1/3, insulin, glucagon and somatostatin. i basally secreted C-peptide, glucagon and ghrelin and released insulin in response either to increasing concentration of glucose or a depolarizing stimulus. The comparison revealed that i beta are remarkably similar to donor derived islets in terms of gene and protein expression profile and similar level of heterogeneity. The ability of i to respond to glucose instead was more related to that of EndoC-beta H1. Discussion: We demonstrated that insulin-producing cells generated from iPSCs recapitulate fundamental gene expression profiles and secretory function of native human beta cells.