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首页> 外文期刊>Journal of biomolecular techniques :JBT. >Comparative Analysis of Non-viral Transfection Methods in Mouse Embryonic Fibroblast Cells
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Comparative Analysis of Non-viral Transfection Methods in Mouse Embryonic Fibroblast Cells

机译:小鼠胚胎成纤维细胞非病毒转染方法的比较分析

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Mouse embryonic fibroblast (MEF) cells are an important in vitro model for developmental biology, disease, and reprogramming studies. However, as with other primary cells, they are challenging to transfect. Although viral gene-delivery methods achieve high gene-delivery efficiency, challenges with cell mutagenesis and safety among others have led to the use and improvement of non-viral gene-delivery methods in MEF cells. Despite the importance of gene delivery in MEF cells, there is limited comparison of method/reagent efficacy. In this study, we compared the effectiveness of different gene-delivery methods and several reagents currently available in MEF cells by introducing a plasmid containing enhanced green fluorescent protein (EGFP). We analyze transfection efficiency by EGFP fluorescence. Our results suggest that two gene-delivery methods—electroporation and magnetofection in combination with a lipid reagent, are the most efficient transfection methods in MEF cells. This study provides a foundation for the selection of transfection methods or reagents when using MEF cells.
机译:小鼠胚胎成纤维细胞(MEF)细胞是用于发育生物学,疾病和重编程研究的重要体外模型。但是,与其他原代细胞一样,它们难以转染。尽管病毒基因递送方法实现了高基因递送效率,但是细胞诱变和安全性等方面的挑战已导致在MEF细胞中使用和改进非病毒基因递送方法。尽管MEF细胞中基因传递的重要性,但方法/试剂功效的比较有限。在这项研究中,我们通过引入包含增强型绿色荧光蛋白(EGFP)的质粒,比较了不同基因递送方法和当前在MEF细胞中可用的几种试剂的有效性。我们通过EGFP荧光分析转染效率。我们的结果表明,两种基因传递方法(电穿孔和磁转染结合脂质试剂)是MEF细胞中最有效的转染方法。该研究为使用MEF细胞时选择转染方法或试剂提供了基础。

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