对定向诱导MSC各分化时期的研究

发布时间：2018-05-19 01:30

本文选题：大鼠 + 骨髓间充质干细胞　；参考：《昆明医学院》2011年硕士论文

【摘要】：[目的]：大鼠骨髓间充质干细胞(rabbit mesenchymal stem cells, MSC)分离纯化培养后,诱导为成骨细胞、心肌细胞和胰岛样细胞,观察不同分化阶段的细胞形态、检测特殊因子或标志物的表达变化。 [方法]：体外分离培养鼠骨髓间充质干细胞,采用文献中的诱导方法诱导,分别于第3、7、14、21天倒置荧光显微镜动态观察细胞形态的变化；并通过RT-PCR技术和酶联免疫吸附技术检测干细胞在诱导不同阶段移植特殊因子或标志物的表达变化。 [结果]：MSC能够正常贴壁成长,抗原表达为CD34-/CD45-/CD73+/CD105+。倒置萤光显微镜观察下细胞形态发生明显变化,RT-PCR检测结果显示GATA-4、Nkx2.5、Sarcomericactin和Connexin-43mRNA向诱导成心肌细胞的过程中表达持续增加,Isulin 1、Isulin 2、CK-19、nestin、Glut2和pdxl mRNA表达在向胰岛样细胞诱导的过程中持续增加,说明在诱导剂诱导下能够向诱导为成骨细胞、心肌细胞和胰岛样细胞分化。 [结论]：①建立了GFP标记的RMSC的体外分离、培养及鉴定的技术平台。②建立了MSC向心肌细胞、成骨细胞和胰岛样细胞分化的诱导体系。⑧RT-PCR结果显示MSC成功向心肌细胞和胰岛样细胞分化,并分泌相应细胞因子。④酶联免疫吸附实验结果显示MSC向胰岛样细胞和成骨细胞分化,并且分泌胰岛素和碱性磷酸酶。
[Abstract]:[objective]: rat bone marrow mesenchymal stem cells (BMSCs) were isolated and cultured in rabbit mesenchymal stem cells, MSC), then induced into osteoblasts, cardiomyocytes and islet like cells. [methods] Rat bone marrow mesenchymal stem cells were isolated and cultured in vitro, induced by the induction method in literature, and the morphological changes of the cells were observed by inverted fluorescence microscope on day 3, 7, 1421, respectively. RT-PCR and enzyme linked immunosorbent assay (Elisa) were used to detect the expression of specific factors or markers in different stages of stem cell transplantation. [results] CD34-/CD45-/CD73 / CD105 was expressed as CD34-/CD45-/CD73 / CD105 antigen. The results of reverse fluorescent microscopy showed that the expression of Glut2 and pdxl mRNA in islet like cells increased continuously during the process of inducing cardiac myocytes by GATA-4Nkx2.5Nkx2.5A, and the expression of Glut2 and pdxl mRNA increased continuously during the induction of cardiac myocytes, and the expression of Glut2 and pdxl mRNA increased during the induction of Glut2 and pdxl mRNA to islet like cells, and the expression of Glut2 and Glut2 in the islet like cells was increased during the induction of Glut2 and pdxl mRNA to islet like cells. These results indicated that induced osteoblasts, cardiomyocytes and islet like cells could be induced. [conclusion] the in vitro isolation, culture and identification of RMSC labeled by GFP were established in vitro. 2. 2. MSC to cardiomyocytes was established by 1: 1. The differentiation of osteoblasts and islet like cells was induced by .8RT-PCR. The results showed that MSC successfully differentiated into cardiomyocytes and islet like cells. The results of enzyme linked immunosorbent assay (Elisa) showed that MSC differentiated into islet like cells and osteoblasts and secreted insulin and alkaline phosphatase.
【学位授予单位】：昆明医学院
【学位级别】：硕士
【学位授予年份】：2011
【分类号】：R329

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