大鼠骨髓基质干细胞诱导分化为神经元样细胞相关分子机制研究

发布时间：2018-06-05 00:17

本文选题：骨髓基质干细胞 + 诱导分化　；参考：《苏州大学》2009年硕士论文

【摘要】： 第一部分还原型谷胱甘肽诱导大鼠骨髓基质干细胞分化过程中NSE、GFAP相关mRNA及蛋白表达变化研究目的:初步探讨还原型谷胱甘肽(GSH)对大鼠骨髓基质干细胞(bone marrow stromalcells,BMSCs)的诱导分化机制。方法:体外培养和纯化大鼠BMSCs,用GSH对其进行诱导,分别于诱导后6h、12h、24h后对其进行实时定量PCR和Westblot检测,测定各组中神经元特异性烯醇化酶(neuron specific enolase,NSE)和胶质纤维酸性蛋白(glial fibrillary acidic protein,GFAP)mRNA及蛋白水平的表达情况。结果:诱导6h、12h、24h后BMSCs分化形成神经元样细胞和胶质细胞样细胞,实时定量PCR和Westblot检测结果表明BMSCs中NSE及GFAP的mRNA与蛋白水平表达明显高于对照组,以12h最为明显。结论:GSH可以诱导BMSCs分化形成神经元样细胞和胶质细胞样细胞,且诱导后NSE及GFAP的mRNA与蛋白水平表达增高。第二部分碱性成纤维细胞生长因子与表皮生长因子诱导大鼠骨髓基质干细胞向神经元样细胞分化相关基因表达谱研究目的:初步探讨大鼠骨髓基质干细胞(BMSCs)向神经元样细胞分化前后基因表达谱的变化,尤其是与神经元发育相关基因的表达情况。方法:体外培养和纯化大鼠BMSCs,用碱性成纤维细胞生长因子(basic fibroblastgrowth factor,bFGF)和表皮生长因子(epidermal growth factor,EGF)对其进行诱导,分别于诱导前和诱导后第七天用Affymetrix大鼠表达谱芯片检测,并且利用生物信息学分析技术比较诱导前后基因表达谱的改变。结果:共有差异基因216条,其中上调基因125条,下调基因91条.与神经元发育密切相关的基因有13条,其中上调7条,下调6条。结论:经体外诱导的BMSCs具有向神经元分化的潜能并能表达一些与神经元分化有关的重要基因。这些基因将是后续研究的重点对象。
[Abstract]:Part I changes in NSE, GFAP related mRNA and protein expression in rat bone marrow stromal cells induced by reduced glutathione
Objective: To explore the mechanism of reduced glutathione (GSH) induced differentiation of bone marrow stromalcells (BMSCs) in rats.
Methods: the rat BMSCs was cultured and purified in vitro, and was induced by GSH. After the induction of 6h, 12h, and 24h, the quantitative PCR and Westblot were measured, and the neuron specific enolase (neuron specific enolase, NSE) and glial fibrillary acidic protein were measured and the protein levels were measured and the protein levels were measured. The expression.
Results: 6h, 12h, and 24h were induced to differentiate into neuron like cells and glial cell like cells. Real-time quantitative PCR and Westblot detection results showed that the expression of NSE and GFAP in BMSCs was significantly higher than that in the control group, and the 12h was the most obvious.
Conclusion: GSH can induce BMSCs to differentiate into neuron like cells and glial cell like cells, and increase the expression of NSE and GFAP mRNA and protein after induction.
Second basic fibroblast growth factors and epidermal growth factor induced gene expression profiles of rat bone marrow stromal cells to neuron like cells
Objective: To investigate the changes in gene expression profiles of rat bone marrow stromal stem cells (BMSCs) before and after differentiation into neuron like cells, especially the expression of genes related to neuron development.
Methods: the rat BMSCs was cultured and purified in vitro. Basic fibroblastgrowth factor (bFGF) and epidermal growth factor (epidermal growth factor, EGF) were used to induce them. The expression chips were detected by Affymetrix rats before and seventh days after induction, and the bioinformatics analysis technique was used. The changes of gene expression profiles before and after induction were compared.
Results: there were 216 different genes, of which 125 were up-regulated and 91 were down regulated. There were 13 genes closely related to the development of neurons, of which 7 were up and 6 were down.
Conclusion: the in vitro induced BMSCs has the potential to differentiate into neurons and can express some important genes related to the differentiation of neurons. These genes will be the focus of follow-up research.
【学位授予单位】：苏州大学
【学位级别】：硕士
【学位授予年份】：2009
【分类号】：R329

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