人骨髓间充质干细胞向胰岛素分泌细胞诱导分化的实验研究

发布时间：2018-05-01 22:08

本文选题：人骨髓间充质干细胞 + 胰岛移植　；参考：《吉林大学》2007年博士论文

【摘要】： 为解决目前胰岛移植根治糖尿病所面临的胰岛来源匿乏和免疫排斥两大难题,本实验以人骨髓间充质干细胞(hMSCs)作为胰岛移植种子细胞,以适宜条件诱导hMSCs分化为胰岛素分泌细胞并初步探讨其分化机制。以密度梯度离心法和贴壁筛选法大量培养扩增hMSCs,利用高糖、尼克酰胺和Exendix-4体外诱导hMSCs向胰岛素分泌细胞分化,通过形态学、酶学、分子生物学和电生理学等方法对诱导后的细胞进行分泌功能和活性鉴定,并以PDX-1和离子通道变化为切入点探讨其分化条件和机制。并利用BrdU标记诱导后细胞移植入糖尿病裸鼠体内,进一步观察其是否具有分泌胰岛素和降低血糖的功能。结果发现:成功地分离培养并扩增hMSCs;体外诱导hMSCs分化为为胰岛素分泌细胞,并且分化后细胞保持良好的增殖活性,其胰岛素的分泌受外环境糖浓度的调节;hMSCs分化机制可能为高糖刺激了hMSCs细胞PDX-1的表达和活化以及Ca2+通道开放,从而激活下游一系列胰岛分化相关基因;BrdU标记诱导后细胞移植入糖尿病裸鼠体内能够存活增殖并具有较好的降低血糖功能,具备了一定的胰岛细胞功能。综上所述,本实验系统全面的探讨了适宜条件下hMSCs分化为胰岛素分泌细胞的可能和机制,为今后更好地利用hMSCs作为胰岛移植的种子细胞治疗胰岛素依赖型糖尿病提供了理论基础和实验依据。
[Abstract]:In order to solve the two major problems of islet origin obscurity and immune rejection faced by islet transplantation to cure diabetes mellitus, human bone marrow mesenchymal stem cells (hMSCs) were used as seed cells for islet transplantation. HMSCs was induced to differentiate into insulin-secreting cells under suitable conditions and its differentiation mechanism was discussed. HMSCs were cultured and amplified by density gradient centrifugation and adherent screening. High glucose, nicotinamide and Exendix-4 were used to induce the differentiation of hMSCs into insulin-secreting cells in vitro. The secretory function and activity of induced cells were identified by molecular biology and electrophysiology, and the differentiation conditions and mechanism of induced cells were discussed with the changes of PDX-1 and ion channels as the starting point. The cells induced by BrdU were transplanted into nude mice to observe whether they could secrete insulin and decrease blood glucose. The results showed that hMSCs were successfully isolated and amplified, hMSCs was induced to differentiate into insulin-secreting cells in vitro, and the cells maintained good proliferative activity after differentiation. Its insulin secretion was regulated by the concentration of glucose in external environment. The mechanism of differentiation of hMSCs might be that high glucose stimulated the expression and activation of PDX-1 and the opening of Ca2 channel in hMSCs cells. As a result of activation of a series of downstream islet differentiation related genes BrdU labeled induced cells transplanted into diabetic nude mice can survive and proliferate and have a better function of lowering blood glucose and have a certain function of islet cells. In conclusion, the mechanism of hMSCs differentiation into insulin-secreting cells under suitable conditions was discussed. It provides a theoretical and experimental basis for the better use of hMSCs as seed cells for islet transplantation in the treatment of insulin-dependent diabetes.
【学位授予单位】：吉林大学
【学位级别】：博士
【学位授予年份】：2007
【分类号】：R329

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