Nanoparticles as transfection agents: a comprehensive study with ten different cell lines
Nanoparticles as transfection agents: a comprehensive study with ten different cell lines
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DOI：
10.1039/c5ra25333k
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发表时间：
2016-01-01
期刊：
影响因子：
3.9
通讯作者：
Epple, Matthias
中科院分区：
文献类型：
--
作者：
Neuhaus, Bernhard;Tosun, Benjamin;Epple, Matthias
转染剂将核酸送入细胞的性能强烈依赖于细胞类型。在一项全面的研究中，将编码增强型绿色荧光蛋白(EGFP)的DNA导入九种不同的细胞系和原代人骨髓间充质干细胞。作为转染剂，使用了两种阳离子多壳层磷酸钙纳米颗粒和市售转染剂脂质体。荧光显微镜下通过计数绿色荧光细胞表达绿色荧光蛋白的百分率和qPCR检测细胞的转染率。此外，通过荧光显微镜测量了荧光磷酸钙纳米颗粒的摄取量。纳米粒和脂质体共同孵育后，用四甲基偶氮唑盐比色法测定细胞存活率。所有类型的细胞都摄取了纳米颗粒(效率不同)，但EGFP的表达差异很大，这表明摄取不一定导致基因的加工。转染率与细胞存活率之间存在明显的相关性，且不依赖于转染剂：高转染率与低细胞存活率明显相关，反之亦然。
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.