体外分步诱导人母体来源胎盘间充质细胞分化为胰岛素分泌细胞

A step-by-step method for differentiating mesenchymal stem cells of maternal origin from human placenta into insulin-producing cells in vitro

目的建立体外诱导人母体来源胎盘间充质细胞分化为胰岛素分泌细胞的分步诱导分化体系。方法分离纯化人母体来源胎盘间充质细胞,体外扩增后进行分步诱导分化,收获的胰岛素分泌细胞分别行免疫细胞化学和ELISA检测,并通过葡萄糖刺激的胰岛素释放试验(GSIS)评价其体外功能。结果分离纯化的人母体来源胎盘间充质细胞经培养及六步诱导分化后最终可形成胰岛素分泌细胞。免疫细胞化学检测诱导后细胞GLP-1、PDX-1和C-Peptide染色为阳性,ELISA结果显示该诱导细胞培养上清中胰岛素有较高的表达。诱导后的胰岛素分泌细胞在低糖和高糖刺激下的胰岛素释放量分别为(20.42±1.58)μIU/mL和(65.1±7.43)μI-U/mL(P<0.01)。结论通过建立体外人母体来源胎盘间充质细胞分化为胰岛素分泌细胞的分步诱导分化体系,获得具有一定胰岛素分泌能力的细胞,为胰岛移植的细胞来源提供了更有效的新途径。

【Objective】 To establish a new step-by-step method for differentiating mesenchymal stem cells of maternal origin from human placenta into insulin-producing cells in vitro.【Methods】 Isolated and purified the mesenchymal stem cells of maternal origin from human placenta,which were differentiated to insulin-producing cells by using a six-step method.The insulin-producing cells were examined by immunocytochemical method and enzyme-linked immuno sorbent assay（Elisa）.The ability of insulin secretion was evaluated through glucose stimulated insulin secretion.【Results】 The isolated and purified mesenchymal stem cells of maternal origin from human placenta could transform to insulin-producing cells by using a six-step method.The results of immunocytochemical method revealed that GLP-1,PDX-1 and C Peptide of the insulin-producing cells were positive.The results of Elisa revealed that the insulin-producing cells also had the expression of insulin.The secretion amount of insulin of the insulin-producing cells was（20.42±1.58） μIU/mL and（65.1±7.43） μIU/mL at low and high concentration of glucose,respectively.【Conclusion】 To establish a new differentiation and induction system can obtain insulin-producing cells that possess insulin secretion ability,which may provide a more effective method to obtain new sources of β cells for islet transplantation.