August Kirchenstein Institute of Microbiology and Virology, Riga Stradins University, Ratsupites 5, 1067 Riga, Latvia.
Medicina (Kaunas). 2012;48(6):324-9.
Liposomal magnetofection is based on the use of superparamagnetic particles and cationic lipids and shows better transfection efficiency than other common nonviral gene delivery methods; however, the distribution of aggregate complexes over the cell surface may be ununiform. The use of a dynamic gradient magnetic field could overcome this limitation. A newly developed device for magnetofection under a dynamic magnetic field was used to compare the transfection efficiency of prostate carcinoma cell line PC3 with that obtained by lipofection and magnetofection.
Reporter plasmid pcDNA3.1LacZ DNA was used in combination with Lipofectamine2000 reagent and superparamagnetic nanoparticles CombiMag. The effects of incubation time under a dynamic magnetic field and a rotation frequency of magnets on transfection efficiency for PC3 cell line were determined. Alternatively, lipofection and liposomal magnetofection were carried out. Transfection efficiency of delivery methods was estimated by β-galactosidase staining; cell viability, by acridine orange/ethidium bromide staining.
Liposomal magnetofection under a dynamic gradient magnetic field demonstrated the highest transfection efficiency: it was greater by almost 21% and 42% in comparison with liposomal magnetofection and lipofection, respectively. The optimal incubation time under dynamic magnetic field and the optimal magnet rotation frequency were 5 minutes and 5 rpm, respectively. Liposomal magnetofection under a dynamic gradient magnetic field was less cytotoxic (7%) than that under a permanent magnetic field (17%) and lipofection (11%).
Our new approach, based on the use of a dynamic gradient magnetic field, enhanced the transfection efficiency and had a less cytotoxic effect on prostate cancer cells in comparison with the standard magnetofection and lipofection.
脂质体磁转染基于超顺磁性颗粒和阳离子脂质的使用,比其他常见的非病毒基因传递方法显示出更好的转染效率;然而,聚集复合物在细胞表面上的分布可能不均匀。使用动态梯度磁场可以克服这一限制。一种新开发的用于动态磁场下磁转染的设备用于比较前列腺癌细胞系 PC3 的转染效率与脂质体转染和磁转染的转染效率。
报告质粒 pcDNA3.1LacZ DNA 与 Lipofectamine2000 试剂和超顺磁性纳米粒子 CombiMag 联合使用。确定在动态磁场下孵育时间和磁体旋转频率对 PC3 细胞系转染效率的影响。或者,进行脂质体转染和脂质体磁转染。通过β-半乳糖苷酶染色估计递送方法的转染效率;通过吖啶橙/溴化乙锭染色估计细胞活力。
动态梯度磁场下的脂质体磁转染显示出最高的转染效率:与脂质体磁转染和脂质体转染相比,分别提高了近 21%和 42%。在动态磁场下的最佳孵育时间和最佳磁体旋转频率分别为 5 分钟和 5rpm。动态梯度磁场下的脂质体磁转染的细胞毒性(7%)低于恒定磁场下的(17%)和脂质体转染的(11%)。
与标准的磁转染和脂质体转染相比,我们基于使用动态梯度磁场的新方法增强了前列腺癌细胞的转染效率,并且对细胞毒性的影响较小。