胎儿骨髓间充质干细胞体外定向诱导分化为胰岛样细胞  

Differentiation of fetal mesenchymal stem cells into pancreatic islet-like clusters in vitro

作　　者：华秀峰[1] 王玮[1] 王海燕[2] 连培文[2] 张守信[2] 陈述林[1] 高伟[1] 李建远[2] 

机构地区：[1]山东省烟台毓璜顶医院内分泌科,山东烟台264000 [2]山东省干细胞工程技术研究中心

出　　处：《中国医师杂志》2006年第10期1297-1299,共3页Journal of Chinese Physician

基　　金：国家863课题基金资助项目(2003AA205008);山东省科学技术发展计划资助项目(021100104)

摘　　要：目的探索体外诱导胎儿骨髓间充质干细胞(MSCs)分化为胰岛样细胞的最佳分化条件。方法取流产胎儿的长骨骨髓,体外培养扩增MSCs,先后予以2-巯基乙醇、表皮生长因子(EGF)、碱性成纤维细胞生长因子(bFGF)、B27及尼克酰胺,通过三阶段诱导MSCs分化为胰岛样细胞,观察细胞形态变化、免疫荧光鉴定巢素蛋白(nestin)、胰十二指肠同源异型盒基因(PDX-1)、胰岛素(insu lin)和胰高糖素(glucagon)的表达、双硫腙染色鉴定胰岛细胞中的锌离子、电化学发光法测定人胰岛素的分泌量。结果MSCs易于体外分离和扩增,诱导二阶段细胞表达nestin及PDX-1,诱导三阶段形成胰岛样细胞团,表达insu lin及glucagon;双硫腙染色呈棕红色;胞浆胰岛素分泌量为81.3±23.6μu/m l;有一定程度葡萄糖刺激的胰岛素释放。结论MSCs可以诱导分化为胰岛样细胞团,尼克酰胺20 mmol/L是合适的促进胰岛细胞分化和成熟的浓度。Objective To study and to optimize culture conditions of islet-like cells induced in vitro from fetus bone marrow（BM） mesenchymal stem cells（MSCs）. Methods BM was obtained from miscarried human fetus. The MSCs between three to eight passages were used to differentiate into islet-like clusters -through three stages of culture supplemented with 2-mercaptoethanol, epidermal growth factor （ EGF）, basic fibroblast growth factor（ bFGF）, B27 and nicotinamide. Results The 2nd stage cells expressed nestin and/or panceatic and duod-enal homeobox 1 （ PDX-1 ）, and the 3rd stage cells formed islet-like clusters expressing insulin and glucagon together with positive dithizone staining. Specific insulin secretion could be detected （81.3±23.6μu/ml） from differentiated MSCs which have the capacity to respond to different glucose concentrations. Conclusion Fetal bone marrow MSCs can be differentiated into pancreatic islet-like clusters, and 20mmol/L nicotinamide could be the optimal concentration in culture.

关 键 词：间质干细胞 骨髓细胞 胰岛 

分 类 号：R329[医药卫生—人体解剖和组织胚胎学]