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为了确定聚乙烯亚胺(polyethlenimine,PEI)转染肺癌A549/DDP细胞的最佳条件,试验以PEI为载体,将含有绿色荧光蛋白的pRNAT-H1.1/Shuttle-GFP表达载体导入肺癌A549/DDP细胞中,在倒置荧光显微镜下观察不同PEI和DNA质量比、细胞密度、血清含量,以及是否去内毒素和线性化重组质粒对聚乙烯亚胺转染肺癌A549/DDP细胞的影响。结果表明:在质量比为2∶1、肺癌A549/DDP细胞于含10%血清中生长至密度为80%时,转染48 h的转染率最高。
To determine the optimal conditions for transfecting lung cancer A549 / DDP cells with polyethleneimine (PEI), pRNAT-H1.1 / Shuttle-GFP expression vector containing green fluorescent protein (EGFP) was transfected into A549 / DDP cells were observed under inverted fluorescence microscope different mass ratio of PEI and DNA, cell density, serum content, and whether to endotoxin and linearized recombinant plasmid polyethyleneimine transfection of lung cancer A549 / DDP cells. The results showed that the transfection efficiency of lung cancer A549 / DDP cells cultured in 10% serum with a density of 80% at the mass ratio of 2:1 was the highest.