大鼠骨髓间充质干细胞与大鼠胰腺岛管上皮细胞间接共培养向胰岛细胞分化的实验研究

Bone marrow mesenchymal stem cells differentiate into insulin producing cells as co-culturing with pancreatic ductal epithelial cells

目的探讨骨髓间充质干细胞(BMSCs)在适当条件下体外分化胰岛素分泌细胞的可能性。方法分离大鼠BMSCs及大鼠胰腺导管上皮细胞分别培养,在大鼠BMSCs培养过程中加入大鼠胰腺导管上皮细胞培养基中的上清部分,双硫腙(DTZ)染色观察诱导后的细胞着色情况,培养7d后进行了反转录-聚合酶链反应(RT-PCR)检测目的基因的表达情况。结果将BMSCs进行分离和纯化,培养至第4代第6天时细胞形态一致,呈"水草"样排列,大鼠胰腺导管上皮细胞表达CK-19;混合培养基诱导的BMSCsDTZ染色阳性,RT-PCR检测有胰腺-十二指肠同源盒基因(PDX-1)及胰岛素(Ins)表达。结论适当的培养环境下,大鼠BMSCs具有向胰岛素分泌细胞分化的能力。

Objective To investigate the feasibility of bone marrow mesenchymal stem cells differentiate into insulin producing cells in appropriate vitro environment. Methods Rat bone marrow mesenchymal stem cells and rat pancreatic ductal epithelial cells were isolated and cultured respectively. Add the supematant medium pancreatic ductai epithelial cells was cultured into the medium The identity of differentiated cells was confirmed by ditbizone staining and analysis of the gene expression for endocrine markers （PDX-1, insulin） by RT-PCR. Results The bone marrow mesenchymal stem cells were isolated and purified. The morph of the cells were coincidence when they were passaged 4 times 6th day and arranged like waterweed. Rat pancreatic dactal epithelial cells expressed CK-19. The bone marrow mesenchymal stein stroma cells were cultured in the mix culture medium. Most cells were stained by DTZ. The cells expressed PDX-1 and Insulin by RT-PCR. Conclusion Rat bone marrow mesenchymal stem ceils have the potentiality to differentiate into insulin producing cells.