人脐带间充质干细胞向胰岛素分泌细胞分化的实验研究

发布时间：2018-02-01 00:03

本文关键词： 脐带间充质干细胞胰岛素分泌细胞分化诱导　出处：《汕头大学》2007年硕士论文　论文类型：学位论文

【摘要】： 目的：探讨人脐带华尔通胶质来源的间充质干细胞(MSCs)分化为胰岛素分泌细胞的可能性，为1型糖尿病的细胞移植治疗提供理论依据和新的细胞来源。材料和方法：从足月剖宫产的新生儿脐带华尔通胶质中分离培养出间充质干细胞(MSCs)，用含尼克酰胺和β巯基乙醇的低糖DMEM培养基(L-DMEM)预诱导后，，以含尼克酰胺和β巯基乙醇的高糖DMEM培养基(H-DMEM)诱导其向胰岛素分泌细胞分化，应用免疫细胞化学检测胰岛素的表达，以双硫腙染色鉴定胰岛β细胞，并用放射免疫法(RIA)检测诱导后细胞培养液中的胰岛素水平，所有检测均以未诱导细胞作为实验对照。结果：从人脐带华尔通胶分离、培养的间充质干细胞，可贴壁生长，体外生长形态类似于成纤维细胞，可以维持未分化状态稳定增殖，体外增殖可超过10代。尼克酰胺和β巯基乙醇联合诱导可使人脐带间充质干细胞(MSCs)向胰岛素分泌细胞分化，分化的细胞表达胰岛β细胞的标记胰岛素(insulin)。双硫腙染色发现脐带华尔通胶来源的间充质干细胞(MSCs)经尼克酰胺和β巯基乙醇诱导后细胞内锌离子增加。通过Insulin放射免疫法(RIA)检测诱导前后细胞培养液中胰岛素的浓度，发现实验组和对照组胰岛素水平分别为6.63mIU／L和3.39mIU／L，差别具有统计学意义(p＜0.05)。结论：人脐带华尔通胶中含有丰富的间充质干细胞(MSCs)，易于培养扩增。人脐带间充质干细胞(MSCs)经尼克酰胺和β巯基乙醇联合诱导后可表达胰岛β细胞的表面标记，可以增加细胞中的锌离子含量使之具备胰岛β细胞的特点，诱导后的细胞以也可以分泌胰岛素。本研究初步证明，用尼克酰胺和β巯基乙醇可以诱导人脐带华尔通胶来源的间充质干细胞(MSCs)成为胰岛素分泌细胞，从而为1型糖尿病细胞移植治疗提供新的细胞来源。
[Abstract]:Objective: To explore the possibility of differentiation of mesenchymal stem cells derived from human umbilical cord Walton glial mesenchymal stem cells into insulin-secreting cells and to provide a theoretical basis and a new cell source for the treatment of type 1 diabetes mellitus. Materials and methods: Mesenchymal stem cells (MSCs) were isolated from the umbilical cord of newborns undergoing caesarean section. L-DMEM was preinduced in low-sugar DMEM medium containing nicotinamide and 尾 -mercaptoethanol. High glucose DMEM medium containing nicotinamide and 尾 -mercaptoethanol was used to induce the differentiation into insulin-secreting cells, and the expression of insulin was detected by immunocytochemistry. Islet 尾 cells were identified by dithizone staining and insulin levels were detected by radioimmunoassay (RIA). Uninduced cells were used as experimental controls. Results: The mesenchymal stem cells isolated from the human umbilical cord and cultured in vitro can grow on the wall, and the growth morphology in vitro is similar to that of fibroblasts, which can maintain stable proliferation in undifferentiated state. The combination of nicotinamide and 尾 -mercaptoethanol could induce the differentiation of human umbilical cord mesenchymal stem cells into insulin-secreting cells. Differentiated cells expressed islet 尾 -labeled insulin insulin.Dithizone staining found mesenchymal stem cells derived from umbilical cord Walton glue mesenchymal stem cells (MSCs). The concentration of insulin in the culture medium was detected by Insulin radioimmunoassay before and after the induction of nicotinamide and 尾 mercaptoethanol. It was found that the insulin levels in the experimental group and the control group were 6.63mIUP / L and 3.39mIUP / L, respectively, and the difference was statistically significant (P < 0.05). Conclusion: There are abundant mesenchymal stem cells (MSCs) in human umbilical cord Walton glue. Human umbilical cord mesenchymal stem cells (MSCs) were induced by nicotinamide and 尾 mercaptoethanol to express the surface markers of islet 尾 cells. It can increase the content of zinc ions in the cells to make them possess the characteristics of islet 尾 cells, and the induced cells can also secrete insulin. Nicotinamide and 尾 -mercaptoethanol can induce mesenchymal stem cells (MSCs) from human umbilical cord warton-glue-derived mesenchymal stem cells (MSCs) to become insulin-secreting cells, thus providing a new cell source for the transplantation of type 1 diabetic cells.
【学位授予单位】：汕头大学
【学位级别】：硕士
【学位授予年份】：2007
【分类号】：R587.1;R329

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