人胚胎多潜能干细胞向胰岛素分泌细胞转化的研究

发布时间：2019-01-10 16:32

【摘要】： 糖尿病在临床上是一种常见病多发病,并发症多,患者的生活质量受严重影响。目前糖尿病的治疗手段有注射胰岛素,胰岛移植,干细胞移植等。常规疗法中注入胰岛素只具有降低血糖的作用,而糖尿病的细胞治疗不止是单纯的降血糖,其根本在于恢复机体调节血糖的功能。干细胞在特定的环境下,具有分化为各种组织、器官的潜能性。因此,利用干细胞诱导分化后的胰岛细胞移植治疗糖尿病,成为一种新的途径。本实验以人胚胎肝脏组织中分离出的多潜能干细胞为研究对象,在高糖环境下,利用bFGF,胰升血糖素样肽和尼克酰胺在体外诱导分化成胰岛素分泌细胞。然后通过形态学检测,RT-PCR,双硫腙染色,免疫荧光染色等对其进行胰岛素分泌功能和活性的检测,从而研究人胚胎多潜能干细胞经诱导分化为胰岛素分泌细胞的潜能和可能性机制。结果显示:在高糖型DMEM培养环境和bFGF、GLP-1、烟酰胺的作用下,人胚胎多潜能干细胞可以分化为胰岛素分泌细胞,RT-PCR可以检测到胰岛素基因、Somatostain和PDX-1的表达,双硫腙染色呈阳性,且胰岛素的分泌受到外环境中葡萄糖浓度的变化。本实验初步探索了人胚胎多潜能干细胞向胰岛素分泌细胞的诱导分化,为糖尿病治疗提供了新的种子细胞来源。
[Abstract]:Diabetes mellitus (DM) is a common disease with many complications, and its quality of life is seriously affected. At present, the treatment of diabetes is insulin injection, islet transplantation, stem cell transplantation and so on. The effect of insulin injection in conventional therapy is only to reduce blood sugar, but the cell therapy of diabetes is not only hypoglycemia, but also restore the function of regulating blood sugar. Stem cells have the potential to differentiate into various tissues and organs in a specific environment. Therefore, transplantation of islet cells induced by stem cells has become a new approach in the treatment of diabetes mellitus. In this study, multipotent stem cells isolated from human embryonic liver tissue were used to differentiate into insulin-secreting cells in vitro using bFGF, glucagon like peptide and nicotinamide. Then the function and activity of insulin secretion were detected by morphological examination, RT-PCR, dithizone staining and immunofluorescence staining. To study the potential and possible mechanism of differentiation of human embryonic pluripotent stem cells into insulin secreting cells. The results showed that human embryonic pluripotent stem cells could differentiate into insulin-secreting cells under high glucose DMEM culture environment and bFGF,GLP-1, nicotinamide, and RT-PCR could detect the expression of insulin gene, Somatostain and PDX-1. Dithizone staining was positive and insulin secretion was affected by glucose concentration in external environment. In this study, the induction and differentiation of human embryonic pluripotent stem cells into insulin-secreting cells were preliminarily explored, which provided a new source of seed cells for the treatment of diabetes.
【学位授予单位】：吉林大学
【学位级别】：硕士
【学位授予年份】：2009
【分类号】：R329

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