ES细胞-D_3诱生的胰岛素分泌细胞及其分泌的胰岛素降糖活性的研究  

Studies of the glucose-reducing activity of insulin-producing cells originated from embryonic stem cells -D3

作　　者：刘星霞[1] 缪兵[2] 李府[1] 马秀峰[1] 时庆[1] 沈柏均[1] 

机构地区：[1]山东大学齐鲁医院低温医学研究室 [2]山东大学医学院生理研究所,山东济南250012

出　　处：《山东大学学报（医学版）》2005年第8期671-673,677,共4页Journal of Shandong University:Health Sciences

基　　金：国家自然科学基金资助课题(30271248)。

摘　　要：目的:探讨ES细胞-D3来源的胰岛素分泌细胞(IPCs)分泌的胰岛素的降糖活性及IPCs移植对糖尿病(DM)鼠的降糖作用。方法:ES细胞-D3培养于经处理的鼠胚成纤维细胞滋养层上保持未分化状态扩增,对数生长期时转入无血清含bFGF的DMEM诱导液进行诱导分化,使其分化为IPCs。于诱导的第21天,用ELISA法检测IPCs受高糖刺激2h后所分泌的胰岛素,同时将分泌的胰岛素静脉注射给小鼠,观察受鼠血糖的变化;将诱生的IPCs移植给链脲菌素(STZ)诱导的DM小鼠,观察受鼠血糖水平的改变。结果:ES细胞-D3体外诱导生成的IPCs所分泌的胰岛素具有降糖活性,且IPCs经两次移植给DM小鼠后第5天,血糖水平较移植前显著降低(P<0.05),第2次移植后第15天,受鼠血糖水平反弹到与移植前没有差别。结论:小鼠ES细胞-D3诱导生成的IPCs能够分泌有活性的胰岛素,且在一段时间内可显著降低DM受鼠的血糖。Objective： To study the glucose-reducing effect of insulin-producing cells （IPCs） derived from embryonic stem cells （ESCs） -D3. Methods： Firstly, ESCs-D3 were induced in serum-free DMEM supplemented with bFGF for more than 2 weeks. Insulin secretion was examined by ELISA, and the function of secreted insulin was determined by glucose-reducing experiment on mice. Secondly, experimental diabetes was induced in 6- to 8-week-old male BALB/C mice by a single intraperitoneal injection （200 mg/kg） of streptozotocin freshly dissolved in 0.1mol/L of citrate buffer, pH 4.5, and the induced IPCs were harvested at day 21 of induction, and then grafted subcutaneously in the shoulder of streptozotocin-diabetic mice to ob- serve its＇ glucose-reducing effect. Results： ESCs-D3 could be induced to differentiate into IPCs in serum-free DMEM supplemented with bFGF. The induced IPCs secreted active insulin, and IPCs transplantation could reduce blood glucose of diabetic mouse significantly. Conclusions： ESCs-D3 can be induced to differentiate into IPCs in serum-free DMEM supplemented with bFGF, and the induced IPCs not only secret active insulin, but also play glucose-reducing roles when engrafted in diabetic mice.

关 键 词：干细胞 胚胎 糖尿病 小鼠 近交BALB C 

分 类 号：R587.1[医药卫生—内分泌]