目的建立更为高效的NOD鼠体外胰岛素分泌细胞诱导体系。方法化学因子分步加入培养基,将大鼠胰岛细胞以半透膜相隔与NOD鼠源性诱导性多能干细胞(NOD-iPSCs)共培养约20d,检测胰岛素分泌细胞的相关功能,并与纯化学因子诱导方法进行对比。结果诱导后成团生长,双硫腙染色呈棕红色,RT-PCR显示胰十二脂肠同源异型盒基因1(PDX-1)从第8天起开始表达,并逐渐增强。免疫荧光染色显示细胞表达胰岛素及C-P,联合组及纯化学因子组诱导培养每隔4d检测至20d,高糖刺激后两组胰岛素分泌均呈逐渐升高趋势。结论 NOD-iPSCs能够在体外诱导生成胰岛素分泌细胞。联合培养体系提供大鼠胰岛细胞功能因子可加速分化,提高效率,可能为一种更高效的诱导方法。 Objective To establish a more efficient in vitro insulin secretion system for NOD mice. METHODS: The pancreatic islet cells were co-cultured with NOD-induced pluripotent stem cells (NOD-iPSCs) with semi-permeable membrane for about 20 days to detect the function of insulin-secreting cells. Chemical factor induction method for comparison. Results The growth of the cells was induced by dithizone staining. The expression of PDX-1 gene was gradually increased from day 8 after RT-PCR. Immunofluorescence staining showed that the cells expressed insulin and C-P, and the combination group and the purified factor group induced the culture to be detected every 4 days to 20 days. After high glucose stimulation, the insulin secretion of both groups increased gradually. Conclusion NOD-iPSCs can induce insulin-producing cells in vitro. Co-culture system to provide rat pancreatic islet cell function factor can accelerate differentiation and improve efficiency, may be a more efficient induction method.