BMP-2、TGF-β3双基因修饰质粒在骨髓间充质干细胞内的表达及对增殖活性的影响
发布时间：2018-11-11 18:25
【摘要】：目的：本实验旨在通过阳离子脂质体包埋BMP-2, TGF-β3双基因修饰质粒转染骨髓间充质干细胞,观察BMP2及TGFβ3在细胞内表达及对细胞增殖活性的影响,为下一步研究双基因表达载体对间充质干细胞成骨分化影响奠定基础。 方法：全贴壁法培养兔骨髓间充质干细胞,进行原代和传代培养,取P2代细胞进行转染,分别将pIRES/BMP-2、 pIRES/TGFp3、 pIRES/BMP2-TGFp3及空质粒转染入干细胞,并设置空白对照组,转染两天后RT-PCR检测各组BMP2及TGFβ3的表达情况,绘制细胞生长曲线,计算细胞倍增时间,比较各组干细胞的增殖情况。 结果：pIRES/BMP2-TGFβ3转染组BMP2及TGFβ3mRNA含量高于单基因组及空白对照组(p0.05),双基因均可获得稳定表达；双基因组细胞倍增时间较单一组及空白对照组明显缩短(p0.05),双基因组具有良好的促进BMCs增殖作用。 结论：双基因修饰质粒转染干细胞可以再细胞内获得稳定的表达,并具有良好的协同促干细胞增殖活性,为进一步研究双基因质粒对诱导间充质干细胞成骨的影响及两者之间的协同效应提供理论基础。
[Abstract]:Aim: to investigate the expression of BMP2 and TGF 尾 3 in bone marrow mesenchymal stem cells (BMSCs) transfected with cationic liposome BMP-2, TGF- 尾 3 modified plasmid, and to observe the effect of BMP2 and TGF 尾 3 on the proliferation of bone marrow mesenchymal stem cells. To lay a foundation for the further study of the effect of double gene expression vector on osteogenic differentiation of mesenchymal stem cells. Methods: rabbit bone marrow mesenchymal stem cells (BMSCs) were cultured by whole adherent method, primary and passage culture, P2 generation cells were transfected, pIRES/BMP-2, pIRES/TGFp3, pIRES/BMP2-TGFp3 and empty plasmids were transfected into stem cells, and blank control group was set up. After two days of transfection, RT-PCR was used to detect the expression of BMP2 and TGF 尾 3 in each group, cell growth curve was drawn, cell doubling time was calculated, and the proliferation of stem cells in each group was compared. Results: the contents of BMP2 and TGF 尾 3mRNA in pIRES/BMP2-TGF 尾 3 transfected group were higher than those in single genome and blank control group (p0. 05). The doubling time of double genome cells was significantly shorter than that of single group and blank control group (p0. 05). The double genome could promote the proliferation of BMCs. Conclusion: transfection of double gene modified plasmids into stem cells can obtain stable expression in the cells and has good synergistic effect on the proliferation of stem cells. It provides a theoretical basis for the further study of the effects of double gene plasmids on the osteogenesis of mesenchymal stem cells and their synergistic effects.
【学位授予单位】：青岛大学
【学位级别】：硕士
【学位授予年份】：2012
【分类号】：R329
本文编号：2325740
[Abstract]:Aim: to investigate the expression of BMP2 and TGF 尾 3 in bone marrow mesenchymal stem cells (BMSCs) transfected with cationic liposome BMP-2, TGF- 尾 3 modified plasmid, and to observe the effect of BMP2 and TGF 尾 3 on the proliferation of bone marrow mesenchymal stem cells. To lay a foundation for the further study of the effect of double gene expression vector on osteogenic differentiation of mesenchymal stem cells. Methods: rabbit bone marrow mesenchymal stem cells (BMSCs) were cultured by whole adherent method, primary and passage culture, P2 generation cells were transfected, pIRES/BMP-2, pIRES/TGFp3, pIRES/BMP2-TGFp3 and empty plasmids were transfected into stem cells, and blank control group was set up. After two days of transfection, RT-PCR was used to detect the expression of BMP2 and TGF 尾 3 in each group, cell growth curve was drawn, cell doubling time was calculated, and the proliferation of stem cells in each group was compared. Results: the contents of BMP2 and TGF 尾 3mRNA in pIRES/BMP2-TGF 尾 3 transfected group were higher than those in single genome and blank control group (p0. 05). The doubling time of double genome cells was significantly shorter than that of single group and blank control group (p0. 05). The double genome could promote the proliferation of BMCs. Conclusion: transfection of double gene modified plasmids into stem cells can obtain stable expression in the cells and has good synergistic effect on the proliferation of stem cells. It provides a theoretical basis for the further study of the effects of double gene plasmids on the osteogenesis of mesenchymal stem cells and their synergistic effects.
【学位授予单位】：青岛大学
【学位级别】：硕士
【学位授予年份】：2012
【分类号】：R329
【参考文献】
相关期刊论文 前1条
1 张恒;田晓滨;孙立;杨述华;胡如印;汪雷;陆延盛;张宇坤;傅德皓;;BMP2、VEGF_(165)双基因共表达质粒在小鼠骨髓基质干细胞的表达[J];贵州医药;2009年05期
，本文编号：2325740
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