抑制消减杂交技术筛选骨髓间充质干细胞诱导分化前后差异表达基因

发布时间：2018-05-27 22:23

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

【摘要】： 骨髓间充质干细胞(BMSCs)具有高度可塑性,在一定诱导条件下具有向成骨细胞、成软骨细胞、肌腱细胞、脂肪细胞以及基质细胞等中胚层细胞分化的能力,最近研究表明中胚层来源的BMSCs还可向外胚层来源的神经元分化。本研究在应用流式细胞术成功分选出大鼠骨髓间充质干细胞的基础上,采用视网膜细胞/骨髓间充质干细胞共培养系统,模拟视网膜体内发育的微环境,诱导骨髓间充质干细胞分化为视网膜神经节样细胞。经免疫组化、神经细胞特异性抗体及流式细胞仪检测,鉴定细胞分化水平;再通过抑制性消减杂交(SSH)技术构建骨髓间充质干细诱导前后的cDNA消减文库,筛选BMSCs向视网膜神经节样细胞分化过程中的差异表达基因。结果显示,利用SSH技术筛选出骨髓间充质干细胞在视网膜细胞诱导前后的差异表达基因GAP43,RGS等,上述研究结果表明BMSCs在诱导后表达某些视网膜神经节细胞所特有的基因,从而为转基因方法诱导BMSCs向视网膜神经节细胞定向分化及其功能性研究奠定实验基础。
[Abstract]:Bone marrow mesenchymal stem cells (BMSCs) have the ability to differentiate into mesodermal cells such as osteoblasts, chondroblasts, tendon cells, adipocytes and stromal cells. Recent studies have shown that BMSCs derived from mesoderm can also differentiate into neurons derived from ectoderm. On the basis of the successful selection of rat bone marrow mesenchymal stem cells by flow cytometry, a retinal / bone marrow mesenchymal stem cell co-culture system was used to simulate the microenvironment of retinal development in vivo. Bone marrow mesenchymal stem cells were induced to differentiate into retinal ganglion like cells. The differentiation level of the cells was identified by immunohistochemistry, neuronal specific antibody and flow cytometry, and the cDNA subtractive library was constructed by suppression subtractive hybridization before and after fine induction of bone marrow mesenchymal stem. To screen the differentially expressed genes during the differentiation of BMSCs into retinal ganglion like cells. The results showed that the differentially expressed genes GAP43 and RGS of bone marrow mesenchymal stem cells (BMSCs) were screened by SSH technique before and after induction. The results showed that BMSCs expressed some specific genes in retinal ganglion cells after induction. The results provide experimental basis for the study of the differentiation and function of BMSCs into retinal ganglion cells by transgenic method.
【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】：R346

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