三种转染试剂介导pGPU6/GFP/Neo真核表达载体转染鸡胚成纤维细胞的比较研究  

Comparison of three different methods in transfection of plasmid pGPU6/GFP/Neo to chicken embryo fibroblasts

作　　者：何星[1] 李华[1] 邵雁[1] 胡莹[1] 顾子春[1] 陈力[1] 马菁晶[1] 兰志勇[1] 

机构地区：[1]浙江大学医学院附属邵逸夫医院整形科,浙江杭州310016

出　　处：《浙江大学学报（医学版）》2013年第6期660-665,共6页Journal of Zhejiang University(Medical Sciences)

基　　金：浙江省自然科学基金资助项目(Y2110413)

摘　　要：目的:采用不同转染试剂介导pGPU6/GFP/Neo质粒转染鸡胚成纤维细胞UMNSAH/DF-1,以获得最优化的方法。方法:分别采用不同剂量(1.25μl、2.0μl、2.5μl)的Lipofectamin2000、Gbfectene-Elite及HilyMax转染试剂介导1μg质粒进行转染,观察其转染效率,测定其转染细胞存活率。结果:HilyMax组的转染效率为(86.85%±2.32%),较Lipofectamin2000组(48.33%±3.24%)、Gbfectene-Elite组(37.35%±5.41%)高(F=18.882,P<0.05);其中,HilyMax 2.5μl组的转染效率最高为(90.53%±1.15%)。Lipofectamin2000组的细胞存活率(65.76%±5.78%)明显低于HilyMax组(89.54%±0.86%)、Gbfectene-Elite组(82.45%±3.56%)(F=90.676,P<0.05)。结论:HilyMax对于鸡胚成纤维细胞具有最好的转染效率和最低的细胞毒性,推荐使用2.5μl HilyMax:1μg质粒进行鸡胚成纤维细胞的转染。Objective： To compare three different methods in transfection of plasmid pGPU6/GFP/ Neo into chicken embryo fibroblast. Methods： Different doses （ 1.25μl, 2μl, 2.5 μl ） of Lipofectamin2000, Gbfectene-Elite and HilyMax were used to transfect 1μg plasmid pGPU6/GFP/Neo. The transfection efficiency was observed by the fluorescence and the cell viabilities were measured. Results： The transfection efficiency of HilyMax was significant higher than that of Lipofectamin2000 and Gbfectene-Elite（86.85% ± 2. 32% vs 48.33% ± 3.24% and vs 37.35% ± 5.41% ; F = 18. 882, P 〈 0.05）. The transfection efficiency of 2. 5μl HilyMax was highest（90. 53% ±1.15% ）. The cell viability of Lipofectamin2000 was significantly lower than that of HilyMax and Gbfectene-Elite（65.76% ± 5.78% vs 89.54% ± 0. 86% and vs 82. 45% ± 3.56% ; F = 90. 676, P 〈 0. 05 ）. Conclusion ： HilyMax has the highest transfection efficiency and the lowest cell toxicity in transfection of plasmid pGPU6/GFP/Neo into chicken embryo fibroblast, with the optimal ratio of 2.5 μl HilyMax ： 1 μg plasmid.

关 键 词：转染 质粒DNA/遗传学 鸡胚成纤维细胞/细胞学 绿色荧光蛋白质类 细胞毒性 

分 类 号：Q782[生物学—分子生物学]