CAJ | 학술논문

利用Lipofectamine 2000(Lipo)介导LacZ及EGFP基因转染人肝星状细胞系LX-2和人肝细胞系Chang Liver,探讨Lipo用量、质粒用量、转染时间、细胞初始接种密度及不同启动子驱动对转染效率的影响.结果显示,两种细胞均能得到高效转染,Chang Liver细胞转染效率(>80%)高于LX-2细胞(>60%).Lipo用量为每孔2 μL,转染后6 h换液,LX-2细胞应用质粒DNA为每孔0.45 μg,Chang Liver细胞应用质粒DNA为每孔0.30～0.45 μg,此时转染效率最高. 巨细胞病毒(CMV)启动子驱动LacZ转染效率与CMV早期增强子/鸡β肌动蛋白(CAG)启动子驱动增强型绿色荧光蛋白(EGFP)转染效率无显著差异;细胞初始接种密度对转染效率无显著影响.
이용Lipofectamine 2000(Lipo)개도LacZ급EGFP기인전염인간성상세포계LX-2화인간세포계Chang Liver,탐토Lipo용량、질립용량、전염시간、세포초시접충밀도급불동계동자구동대전염효솔적영향.결과현시,량충세포균능득도고효전염,Chang Liver세포전염효솔(>80%)고우LX-2세포(>60%).Lipo용량위매공2 μL,전염후6 h환액,LX-2세포응용질립DNA위매공0.45 μg,Chang Liver세포응용질립DNA위매공0.30～0.45 μg,차시전염효솔최고. 거세포병독(CMV)계동자구동LacZ전염효솔여CMV조기증강자/계β기동단백(CAG)계동자구동증강형록색형광단백(EGFP)전염효솔무현저차이;세포초시접충밀도대전염효솔무현저영향.
PAAV- CMV-LacZ plasmid and pAAV-CAG-EGFP plasmid were transfected into a kind of human hepatic stellate cell line-LX-2 and a kind of human hepatocyte line-Chang Liver using Lipofectamine 2000, respectively. The impacts of liposome and plasmid amount, transfection time course, initial plated density and different promoter used on transfection efficiency were discussed. It indicated that both kinds of cell lines were highly transfectable, and the transfection efficiency of Chang Liver cells (>80%) was higher than LX-2 cells (>60%). The transfection efficiency turned out to be the highest while using 2 μL·well~(-1) of liposome, 0.45 μg·well~(-1) of plasmid DNA for LX-2 cells and 0.30～0.45 μg·well~(-1) of plasmid DNA for Chang Liver cells after 6 hours of transfection. There was no significantly difference in transfection efficiency whether the gene was driven by CMV or CAG promoter. The initial plated density seemed to have no obvious influence on the transfection efficiency, either.