多途径将骨髓间充质干细胞制备成类β细胞的体外实验研究

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...

Experimental study on multi-ways preparations of beta-like cells from the bone-marrow mesenchymal stem cells in vitro

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,NeuroD1 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 stimulation 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.