Availability of an efficient transfection protocol is the first determinant in success of gene transferring studies in mammalian cells which is accomplished experimentally for every single cell type. Herein, we provide data of a comparative study on optimization of transfection condition by electroporation and chemical methods for Huh-7 and Vero cells. Different cell confluencies, DNA/reagent ratios and total transfection volumes were optimized for two chemical reagents including jetPEI™ and Lipofectamine™ 2000. Besides, the effects of electric field strength and pulse length were investigated to improve electroporation efficiency. Transfection of cells by pEGFP-N1 vector and tracking the expression of GFP by FACS and Fluorescence Microscopy analysis were the employed methods to evaluate transfection efficiencies. Optimized electroporation protocols yielded 63.73 ± 2.36 and 73.9 ± 1.6% of transfection in Huh-7 and Vero cells respectively, while maximum achieved level of transfection by jetPEI™ was 14.2 ± 0.69 and 28 ± 1.11% Huh-7 and Vero cells, respectively. Post transfectional chilling of the cells did not improve electrotransfection efficiency of Huh-7 cells. Compared to chemical based reagents, electroporation showed superior levels of transfection in both cell lines. The presented protocols should satisfy most of the experimental applications requiring high transfection efficiencies of these two cell lines.

中文翻译：

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Huh-7 和 Vero 细胞转染方法的优化：比较研究。


有效转染方案的可用性是哺乳动物细胞基因转移研究成功的首要决定因素，该研究是针对每种细胞类型通过实验完成的。在此，我们提供了 Huh-7 和 Vero 细胞电穿孔和化学方法优化转染条件的比较研究数据。针对jetPEI™和Lipofectamine™ 2000等两种化学试剂，优化了不同的细胞融合度、DNA/试剂比例和总转染体积。此外，还研究了电场强度和脉冲长度的影响，以提高电穿孔效率。采用pEGFP-N1载体转染细胞并通过FACS和荧光显微镜分析追踪GFP的表达是评估转染效率的方法。优化的电穿孔方案在 Huh-7 和 Vero 细胞中分别产生 63.73 ± 2.36 和 73.9 ± 1.6% 的转染率，而 jetPEI™ 在 Huh-7 和 Vero 细胞中分别达到的最大转染水平分别为 14.2 ± 0.69 和 28 ± 1.11%。细胞转染后冷冻并没有提高 Huh-7 细胞的电转染效率。与基于化学的试剂相比，电穿孔在两种细胞系中均显示出更高水平的转染。所提出的方案应满足大多数需要这两种细胞系高转染效率的实验应用。