Zhi De-fu, Wang Bing, Cui Shao-hui, Yang Bao-ling, Zhao Bu-diao, Zhao Yi-nan, Jiang Yun-xia, Yu Shi-jun, Zhang Shu-biao
College of Chemistry and Chemical Engineering, Liaoning Normal University, Dalian 116029, China.
Yao Xue Xue Bao. 2009 May;44(5):553-7.
In order to study the important factors involved in cationic liposome-mediated gene transfer, Lipofectamine 2000 or DOTAP was evaluated using three types of cells (Hep-2, MCF-7 and SW-480) in vitro transfection efficiencies. Different properties of the two reagents were analyzed and compared by DNA arrearage assay and MTT assay. Both Lipofectamine 2000 and DOTAP had strong capability to combine with DNA; Lipofectamine 2000 can get higher transfection efficiency of the three cells by using GFP as report gene, meanwhile, DOTAP can also get higher transfection efficiency against Hep-2 cell. However, DOTAP showed lower transfection efficiency against MCF-7 and SW-480 cell. On the other hand, the cytotoxicity assay showed that over 85% cell viability of MCF-7 cell could be achieved both by Lipofectamine 2000 and DOTAP under the optimal transfection condition. Relatively speaking, Lipofectamine 2000 has very high transfection efficiency in a broad range of cell lines, but because of the special selectivity of cell type on liposome, DOTAP also has a broad application prospect.
为了研究阳离子脂质体介导的基因转移所涉及的重要因素,使用三种细胞(Hep-2、MCF-7和SW-480)对Lipofectamine 2000或DOTAP进行体外转染效率评估。通过DNA滞留试验和MTT试验分析并比较了这两种试剂的不同特性。Lipofectamine 2000和DOTAP都具有很强的与DNA结合的能力;以绿色荧光蛋白(GFP)作为报告基因时,Lipofectamine 2000对这三种细胞能获得较高的转染效率,同时,DOTAP对Hep-2细胞也能获得较高的转染效率。然而,DOTAP对MCF-7和SW-480细胞的转染效率较低。另一方面,细胞毒性试验表明,在最佳转染条件下,Lipofectamine 2000和DOTAP均可使MCF-7细胞的细胞活力超过85%。相对而言,Lipofectamine 2000在广泛的细胞系中具有非常高的转染效率,但由于脂质体对细胞类型有特殊的选择性,DOTAP也具有广阔的应用前景。