【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。更多还原

【Abstract】 Objective: To compare three different methods in transfection of plasmid pGPU6/GFP/ Neo into chicken embryo fibroblast. Methods: Different doses( 1. 25 μl,2 μl,2. 5 μl) of Lipofectamin2000,Gbfectene-Elite and HilyMax were used to transfect 1 μg 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. 5 μl 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.更多还原