重组绿色荧光蛋白腺相关病毒载体转染293细胞和CD34^＋细胞

目的研究重组绿色荧光蛋白腺相关病毒载体分别转染293细胞(人胚肾细胞)和人CD34^ 细胞后绿色荧光蛋白的表达情况。方法用CS-3000血细胞分离机分离骨髓单个核细胞，免疫磁性分离仪纯化CD34^ 细胞，重组绿色荧光蛋白腺相关病毒载体分别转染两种细胞，并通过荧光显微镜、流式细胞仪对绿色荧光蛋白的表达进行分析。结果荧光显微镜下两种细胞均可见绿色荧光蛋白表达，流式细胞仪于一定时间内检测293细胞的基因转导效率最高为32．8％，CD34^ 细胞的基因转导效率最高为25％．在所观察的时间内，转染率呈先升高后降低趋势。结论重组绿色荧光蛋白腺相关病毒载体能转染293和CD34^ 细胞，绿色荧光蛋白基因可作为良好的报告基因，为将来的干细胞基因治疗打下良好基础。

Transfection of human 293 and CD34+ cells with recombinant green fluorescent protein adeno- associated virus]

OBJECTIVE: To investigate the expressions of recombinant green fluorescent protein (GFP) in 293 cells (human embryonic kidney cells) and CD34(+) cells transfected with adeno-associated virus carrying the gene encoding the recombinant GFP. METHOD: Mononuclear cells from the bone marrow were collected on cell separator CS-3000, and CD34(+) cells were separated on immunomagnetic MiniMACS columns. CD34(+) cells and 293 cells were transfected with recombined adeno-associated virus respectively, and the expressions of the GFP were detected by flow cytometry and fluorescence microscopy. RESULT: GFP expression was observed in the two cells under fluorescent microscope. The highest transfection efficiency of the recombined adeno-associated virus was about 32.8% in 293 cells and 25% in CD34(+) cells, and decreased with the prolongation of transfection time. CONCLUSION: Adeno-associated viral vector encoding recombinant GFP can be transfected into human 293 cells and CD34(+) cells, which provides a basis for future gene therapy.