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目的探讨采用脂质体介导的基因转移技术将重组pEGFP-N1/Rapsyn质粒转染至小鼠骨髓间充质干细胞(BMSCs)的可能性,并观察Rapsyn蛋白在靶细胞中的表达。方法采用全骨髓贴壁培养法分离纯化C57BL/6小鼠的BMSCs,在体外进行扩增、传代。取第3代细胞,采用脂质体介导的基因转移技术将重组pEGFP-N1/Rapsyn质粒转染至BMSCs中,并置荧光显微镜下观察转染结果;采用Western blot法检测靶细胞中Rapsyn蛋白的表达。结果 BMSCs经转染后24h可在荧光显微镜下观察到绿色荧光,转染pEGFP-N1/Rapsyn质粒的BMSCs可稳定表达Rapsyn蛋白。结论利用脂质体介导法可将Rapsyn基因转染至BMSCs中,并能稳定表达Rapsyn蛋白。
Objective To investigate the possibility of transfection of recombinant pEGFP-N1 / Rapsyn plasmid into mouse bone marrow mesenchymal stem cells (BMSCs) by liposome-mediated gene transfer and to observe the expression of Rapsyn protein in target cells. Methods BMSCs of C57BL / 6 mice were isolated and purified by whole bone marrow adherent culture and amplified and passaged in vitro. The third generation of cells were transfected with recombinant pEGFP-N1 / Rapsyn plasmid into BMSCs by liposome-mediated gene transfer, and the transfection results were observed under a fluorescence microscope; Rapsyn protein was detected by Western blot expression. Results BMSCs transfected with pEGFP-N1 / Rapsyn plasmid could stably express Rapsyn protein 24h after transfection. Conclusion The Rapsyn gene can be transfected into BMSCs by liposome-mediated method and the Rapsyn protein can be stably expressed.