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目的探索人脐带间充质干细胞(hUCMSC)向胰岛素分泌细胞(IPCs)分化的高效、定向诱导方案。方法在传统诱导方案基础上,加入胰岛新生相关蛋白肽,诱导hUCMSCs分化为IPCs,并通过形态学观察、流式细胞术、荧光定量PCR、免疫荧光、ELISA鉴定IPCs。结果改良组IPCs的C肽阳性细胞率较传统组有明显提高(P<0.05),胰腺发育相关转录因子PDX-1、Ngn3、NeuroD1、MafA、insulin、Glut-2 mRNA表达明显提高(P<0.05)。同时,改良组蛋白C肽、PDX-1、Nkx6.1表达阳性,葡萄糖刺激试验证实胰岛素及C肽分泌能力也明显增强(P<0.05)。结论胰岛新生相关蛋白肽可促进hUCMSCs分化为IPCs,但仍未达到正常人胰岛β细胞功能。
Objective To explore the efficient and targeted induction of hUCMSCs differentiation into insulin-secreting cells (IPCs). Methods Based on the traditional induction protocol, pancreatic islet-derived peptide was added to induce hUCMSCs to differentiate into IPCs. IPCs were identified by morphological observation, flow cytometry, real-time PCR, immunofluorescence and ELISA. Results The C-peptide positive rate of IPCs in the modified group was significantly higher than that in the traditional group (P <0.05), and the expressions of PDX-1, Ngn3, NeuroD1, MafA, Glut- ). At the same time, the expression of PDX-1 and Nkx6.1 was improved, while the secretion of insulin and C-peptide was also significantly enhanced by glucose stimulation test (P <0.05). Conclusion Islet-associated protein peptide can promote the differentiation of hUCMSCs into IPCs, but still not achieve the normal human pancreatic β-cell function.