重组腺病毒介导β半乳糖苷酶基因转染内皮祖细胞的实验研究

The Experimental Studies of Recombinant Adenovirus Vector-mediated β-Galactosidase Gene Transfected to Endothelial Progenitor Cells

目的测定重组腺病毒介导β半乳糖苷酶基因对内皮祖细胞的转染率,为利用人类单纯疱疹病毒胸苷激酶(HSV-TK)基因转染内皮祖细胞治疗肺癌提供依据。方法将转染β半乳糖苷酶(LacZ)的重组腺病毒AD5F35(AD5F35LacZ)加入培养有内皮祖细胞的24孔板,转染内皮祖细胞。转染后用LacZ检测试剂盒染色,计算转染率。结果在光学显微镜下观察,蓝色细胞为AD5F35LacZ基因转染成功的内皮祖细胞。在不同感染复数MOI的前提下,腺病毒对内皮祖细胞的转染率是不同的,在一定的范围内,转染率随MOI的增大而增大,在MOI达到400时,转染率最大,为98.38%±1.25%。结论重组腺病毒可成功转染内皮祖细胞,转染率随MOI的增大而增大,当感染复数为400时转染率最大。

Objective To determine the transfection efficiency of recombinant adenovirus to endothelial progenitor cells（EPCs） and provide the base of lung cancer therapy by transfecting human herpes simplex virus-thymidine kinase （HSV-TK） gene to EPCs. Methods Admove recombinant adenovirus 5F35（AD5F35） which transfected with β- galactosidase（AD5F35LacZ） to the 24 well plate cultivated with EPCs and transfect the EPCs. Stain the EPCs with LacZ kit and calculate the transfection efficiency. Results The blue stain cells were cells transfected successfully with AD5F35LacZ under the optical microscope. The transfection efficiencies of adenovirus to EPCs were different under the premise of the different multiplicity of infection（MOI）. In a certain range, the transfection efficiencies rise with the MOI rise. When MOI was 400,the proportion of blue stain cell is the highest, which was 98.38±1.25M. Conclusion Recombinant adenovirus can transfect EPCs successfully. The transfection efficiencies rise with the MOI rise. When the MOI is 400,the transfection efficiency is the highest.