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猪肾细胞脂质体磁转染系统的构建与表征

[Construction and characterization of liposomal magnetofection system in pig kidney cells].

作者信息

Chen Wenjie, Cui Haixin, Zhao Xiang, Cui Jinhui, Wang Yan, Sun Changjiao

出版信息

Sheng Wu Gong Cheng Xue Bao. 2014 Jun;30(6):972-81.

Abstract

Magnetic nano gene vector is one of the non-viral gene vectors, modified by functional group to bind cationic transfect reagents. Coupling magnetofection with the universal lipofection we developed a novel somatic cell transfection method as the so-called liposomal magnetofection (LMF). This approach is potential to provide somatic cell cloning with stable genetic cell lines to cultivate transgenic animals. In order to construct such liposomal magnetic gene vectors complexes system, we used nano magnetic gene vector to combine with liposomal cationic transfect reagents by molecular self-assembly. This vectors system successfully carried exogenous gene and then transfected animal somatic cells. Here, we conducted atomic force microscopy (AFM), zeta potential-diameter analysis and other characterization experiments to investegate the size distribution and morphology of magnetic nanoparticles, the way of the vectors to load and concentrate DNA molecules. Our data reveal that, the LMF of Pig Kidney cells exhibited higher transfection efficiency comparing with the transfection mediated by the commercial lipofectamine2000. Moreover, LMF method overcomes the constraint of transient expression mediated by lipofection. Meanwhile, MTT assay showed low cytotoxicity of LMF. Hence, LMF is a feasible, low cytotoxic and effective method of cell transfection.

摘要

磁性纳米基因载体是非病毒基因载体之一,通过功能基团修饰以结合阳离子转染试剂。将磁转染与我们开发的通用脂质体转染相结合,我们开发了一种新型的体细胞转染方法,即所谓的脂质体磁转染(LMF)。这种方法有可能为体细胞克隆提供稳定的遗传细胞系,以培育转基因动物。为了构建这样的脂质体磁性基因载体复合物系统,我们通过分子自组装将纳米磁性基因载体与脂质体阳离子转染试剂相结合。该载体系统成功携带外源基因并转染动物体细胞。在此,我们进行了原子力显微镜(AFM)、zeta电位-直径分析等表征实验,以研究磁性纳米颗粒的尺寸分布和形态、载体加载和浓缩DNA分子的方式。我们的数据表明,猪肾细胞的脂质体磁转染与市售脂质体转染试剂Lipofectamine2000介导的转染相比,具有更高的转染效率。此外,脂质体磁转染方法克服了脂质体介导的瞬时表达的限制。同时,MTT分析显示脂质体磁转染的细胞毒性较低。因此,脂质体磁转染是一种可行、低细胞毒性且有效的细胞转染方法。

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