诱导型多能干细胞体外向胰岛素分泌细胞分化及对血糖的调控

Insulin-secreting cells from induced pluripotent stem cells regulate blood glucose levels in vitro

BACKGROUND:Mouse pluripotent stem cells are induced to differentiate into insulin-secreting cells that can effectively improve blood glucose levels in diabetic mice. OBJECTIVE:To detect mRNA and protein levels of insulin-like cell clusters from induced pluripotent stem cells and to investigate the function of insulin-secreting cells in vitro and in vivo.METHODS:Mouse induced pluripotent stem cells cultured in vitro were induced to differentiate into insulin-secreting cells using combined inducers through three stages. The morphology of endodermal cells, islet-derived progenitor cells and mature islet cells in each stage was observed and relative gene expression levels were detected by PCR. Mature insulin-like cell clusters underwent dithizone staining and functions of insulin released in vitro were observed by ELISA assay. Finally, the insulin-secreting cells were transplanted into the subrenal capsule of diabetic mice, and then blood glucose levels were observed. RESULTS AND CONCLUSION:The mature spherical insulin-like cell clusters were successfully obtained in vitro, which were in iron red by dithizone staining, and expression of insulin mRNA was determined by PCR. The insulin-like cell clusters could secrete insulin in response to various blood glucose levels by ELISA assay. In addition, after the cells clusters were transplanted into the subrenal capsule of mice with type 1 diabetes, the blood glucose levels were marbedly improved.