目的 通过诱导法和转基因法将小鼠骨髓间充质干细胞(BMSCs)制备成类β细胞.方法 在体外分离、培养小鼠BMSCs,运用细胞因子诱导剂诱导和共培养诱导,将BMSCs 诱导分化为类β细胞,通过逆转录病毒载体将外源性胰岛素基因转入小鼠BMSCs,使其表达外源性胰岛素基因;获得分泌胰岛素的类β细胞;通过腺病毒载体将PDX-1、NeuroD1 及MafA 基因转导入小鼠BMSCs,刺激内源性的胰岛素基因的表达,制备类β细胞,比较四种途径制备类β细胞的效果.结果 四种方法均能诱导出类β细胞,均有胰岛素分泌,但诱导法和转三基因法制备出的类β细胞产量均较低,转胰岛素基因制备的类β细胞效率高,但对葡萄糖刺激无反应.结论 多细胞因子诱导法和转三基因法制备的类β细胞在功能上更加接近β细胞,但需提高制备效率;转胰岛素基因法制备的类β细胞的功能有待进一步提高.%Objective To prepare beta-like cells from BMSCs by induction and transgene. Methods The bone marrow mesenchymal stem cells of mouse were separated and cultuted in vitro. The multi-cell factors induction and co-cultured induction were used to transform the BMSCs to beta-like cells. Exogenetic insulin gene was transfected to mBMSCs by a recombinant retrovirus MSCV and expressed in the cells, then, PDX-1, NeuroDl and the MafA gene were transfected to BMSCs by the adenovirus. The endogenous insulin gene was stimulated to express, then the beta-like cells were obtained. The efficiency of the four ways was compared each other. Results The beta-like cells were gained from four kinds of methods and could secret the insulin. The beta-like cells induced by multi-cell factors or three-transgenes had the response to the glucose stimula-tion insulin secretion, but the outputs of two methods were low. The efficiency of insulin-transgene way was high, but this kind of beta-like cells did not response to the glucose stimulation. Conclusion The beta-like cells prepared by multi-cell factor induction or three-transgenes are more like beta cells in the function, but the efficiency of production must be raised. The beta-like cells' function of insulin-transgene way needs to improve.
展开▼
