小鼠胚胎干细胞体外诱导分化为胰岛素分泌细胞  

Induced differentiation of mouse embryonic stem cells into insulin-secreting cells in vitro

作　　者：邓爱民[1] 林戈[2] 卢光琇[2] 

机构地区：[1]江门市中心医院生殖中心,广东江门529000 [2]中南大学生殖与干细胞工程研究所,湖南长沙410078

出　　处：《基础医学与临床》2009年第6期575-579,共5页Basic and Clinical Medicine

基　　金：863基金(200AA02A102)

摘　　要：目的建立小鼠胚胎干细胞(mESCs)分化为胰岛素分泌细胞的体外诱导体系。方法mESCs经拟胚体(EB)阶段,筛选出nestin阳性细胞,扩增培养后,用不同浓度烟碱对nestin阳性细胞进行诱导,观察诱导细胞的形态变化,并进行免疫组化染色、流式细胞仪分析和胰岛素分泌实验。结果不同大小EB中nestin阳性细胞比例不同,直径为100μm大小EB中较多。nestin阳性细胞经烟碱诱导15d后,可形成分泌胰岛素的胰岛样细胞团,大部分细胞团胰岛素抗体检测为阳性。以10mmol/L烟碱诱导分化后得到的胰岛素分泌细胞较0mmol/L和5mmol/L多,这些细胞在不同浓度葡萄糖刺激下能分泌不同量胰岛素。结论10mmol/L烟碱诱导小鼠ESCs15d,可获得较多胰岛素分泌细胞。Objective To establish a system in which mouse embryonic stem cells （mESCs） differentiate into insulin-secreting cells in vitro. Methods mESCs were cultured to form embryoid bodies（EBs）. EBs were cultured in serum-free medium to obtain nestin-positive cells. The selected nestin-positive cells were expanded, then added nicotinamide into the medium to induce the differentiation of nestin-positive cells. Immunochemistry staining and flow cytometry analysis were made on 15th day after induction. Results Flow cytometry analysis revealed that the percentage of nestin-positive cells were different from EBs. The percentage of nestin-positive cells from EBs were higher than those of other diameters. Nestin-positive cells induced by nicotinamide formed islet-like cell clusters. Percentage of insulin-positive cell induced by 10 mmol/L nicotinamide was higher than those induced by 0 mmol/L or 5 mmol/L nicotinamide （P 〈 0. 01 ）. The induced cells secreted insulin as respond to glucose. Conclusion More insulin-secreting cells can be obtained from the nestin-positive ceils through treatment of 10 mmol/L nicotinamide for 15 days.

关 键 词：胚胎干细胞 分化 胰岛素 I型糖尿病 

分 类 号：Q813.11[生物学—生物工程]