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Nanoparticles as transfection agents: a comprehensive study with ten different cell lines

机译:纳米粒子作为转染剂:用十种不同的细胞系综合研究

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摘要

The performance of transfection agents to deliver nucleic acids into cells strongly depends on the cell type. In a comprehensive study, nine different cell lines and primary human mesenchymal stem cells were transfected with DNA encoding for enhanced green fluorescent protein (eGFP). As transfection agents, two kinds of cationic multi-shell calcium phosphate nanoparticles and the commercially available transfection agent Lipofectamine were used. The transfection efficiency was measured by fluorescence microscopy by counting the percentage of green fluorescent cells which expressed eGFP as well as qPCR. Furthermore, the uptake of fluorescent calcium phosphate nanoparticles was measured by fluorescence microscopy. The cell viability was measured by the MTT test after incubation with nanoparticles and Lipofectamine. All cell types took up nanoparticles (with different efficiency), but the expression of eGFP was strongly different, demonstrating that the uptake not necessarily leads to processing of a gene. A clear correlation was found between the transfection efficiency and the cell viability that was independent on the transfection agent: a high transfection efficiency was clearly correlated with a low cell viability and vice versa.
机译:转染试剂的性能将核酸递送到细胞中强烈取决于细胞类型。在综合研究中,将九种不同的细胞系和初级人间充质干细胞用编码的DNA编码转染用于增强的绿色荧光蛋白(EGFP)。作为转染剂,使用两种阳离子多壳磷酸钙纳米粒子和市售的转染剂脂质溶剂。通过计算表达EGFP的绿色荧光细胞以及QPCR的绿色荧光细胞的百分比,通过荧光显微镜测量转染效率。此外,通过荧光显微镜测量荧光磷酸钙纳米粒子的摄取。通过与纳米颗粒和Lipofectamine温育后MTT试验测量细胞活力。所有细胞类型占纳米颗粒(具有不同的效率),但EGFP的表达强烈不同,表明摄取不一定导致基因的加工。在转染剂上的转染效率和细胞活力之间发现了明确的相关性:高转染效率与低细胞活力明显相关,反之亦然。

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  • 来源
    《RSC Advances》 |2016年第22期|共11页
  • 作者单位

    Univ Duisburg Essen Inorgan Chem &

    Ctr Nanointegrat Duisburg Essen Ce Univ Str 5-7 D-45117 Essen Germany;

    Univ Duisburg Essen Inorgan Chem &

    Ctr Nanointegrat Duisburg Essen Ce Univ Str 5-7 D-45117 Essen Germany;

    Univ Duisburg Essen Inorgan Chem &

    Ctr Nanointegrat Duisburg Essen Ce Univ Str 5-7 D-45117 Essen Germany;

    Univ Duisburg Essen Univ Hosp Essen Inst Med Microbiol Essen Germany;

    Univ Duisburg Essen Univ Hosp Essen Inst Med Microbiol Essen Germany;

    Univ Duisburg Essen Inorgan Chem &

    Ctr Nanointegrat Duisburg Essen Ce Univ Str 5-7 D-45117 Essen Germany;

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  • 原文格式 PDF
  • 正文语种 eng
  • 中图分类 化学;
  • 关键词

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