胰腺导管上皮细胞来源的胰岛样细胞移植治疗1型糖尿病

目的 观察小鼠胰腺导管上皮细胞向胰岛样细胞转分化及其在治疗糖尿病小鼠中的作用.方法 分离培养昆明小鼠胰腺导管上皮细胞,经体外扩增及诱导培养后,进行体外和体内功能评价.结果 小鼠胰腺导管上皮细胞经体外扩增和诱导分化后,逆转录-聚合酶链反应(RT-PCR)显示培养1周和2周后胰岛素mRNA的IA值为(1.892±0.119比3.135±0.092,P＜0.05),胰高血糖素为(1.564±0.087比2.271±0.042,P＜0.05),表达明显上调;胰岛样细胞对高糖刺激(15.0 mmol/L)的胰岛素释放较低糖(5.6 mmol/L)时增加了1.7倍(52.3±10.5)mmol/L比(30.2±9.7)mmol/L,P＜0.05];DTZ染色阳性;将其移植入糖尿病小鼠体内后,移植组小鼠较对照组血糖下降,差异有统计学意义(P＜0.05).结论 昆明小鼠胰腺导管上皮细胞在体外培养条件可转分化胰岛素分泌细胞,该细胞团可在体内环境下发挥生物学功能.

Abstract：

Objective To investigate the differentiation of stem cells deived from mouse pancreatic ductal epithelial cells toward insulin secreting cells and the potential application for diabetes therapy.Methods Pancreatic ductal epithelial cells were separated and differentiated into islet-like cells. Study was performed to determine whether these islet-like clusters could secrete insulin in response to glucose both in vivo and in vitro. Results Pancreatic ductal epithelial cells were separated and cultured in vitro.The expression of insulin mRNA in the stem cells was significantly up-regulated ( 1. 892 ±0. 119 vs 3. 135± 0. 092,P ＜ 0. 05 ), and also in glucagon ( 1. 564 ± 0. 087 vs 2. 271 ± 0. 042, P ＜ 0. 05 ). Immunofluoresence staining indicated that there were a lot of insulin positive cells. Insulin released from cells in response to glucose stimulation in vitro was increased ( 52. 3 ± 10. 5 ) mmol/L vs ( 30. 2± 9. 7 ) mmol/L, P ＜0. 05 ]. Hyperglycemia in diabetic animals was alleviated after cell transplantation. Conclusion Islet-like cell clusters generated in vitro from Kunming mice pancreatic ductal epithelial cells could secrete insulin and have some effects on reversing the diabetes in diabetic mice.

Islet-like cells differentiated from murine pancreatic ductal epithelial cells implanted to diabetic mice

Objective To investigate the differentiation of stem cells deived from mouse pancreatic ductal epithelial cells toward insulin secreting cells and the potential application for diabetes therapy.Methods Pancreatic ductal epithelial cells were separated and differentiated into islet-like cells. Study was performed to determine whether these islet-like clusters could secrete insulin in response to glucose both in vivo and in vitro. Results Pancreatic ductal epithelial cells were separated and cultured in vitro.The expression of insulin mRNA in the stem cells was significantly up-regulated ( 1. 892 ±0. 119 vs 3. 135± 0. 092,P ＜ 0. 05 ), and also in glucagon ( 1. 564 ± 0. 087 vs 2. 271 ± 0. 042, P ＜ 0. 05 ). Immunofluoresence staining indicated that there were a lot of insulin positive cells. Insulin released from cells in response to glucose stimulation in vitro was increased ( 52. 3 ± 10. 5 ) mmol/L vs ( 30. 2± 9. 7 ) mmol/L, P ＜0. 05 ]. Hyperglycemia in diabetic animals was alleviated after cell transplantation. Conclusion Islet-like cell clusters generated in vitro from Kunming mice pancreatic ductal epithelial cells could secrete insulin and have some effects on reversing the diabetes in diabetic mice.