Chang Chun-Chi, Mao Mao, Liu Yang, Wu Mina, Vo-Dinh Tuan, Yuan Fan
Department of Biomedical Engineering, Duke University, Durham, North Carolina, USA.
Department of Biomedical Engineering, Duke University, Durham, North Carolina, USA. ; Department of Chemistry, Duke University, Durham, North Carolina, USA.
Cell Mol Bioeng. 2016 Dec;9(4):538-545. doi: 10.1007/s12195-016-0452-9. Epub 2016 Jun 13.
Electrotransfection has been widely used as a versatile, non-viral method for gene delivery. However, electrotransfection efficiency (eTE) is still low and unstable, compared to viral methods. To understand potential mechanisms of the unstable eTE, we investigated effects of electrode materials on eTE and viability of mammalian cells. Data from the study showed that commonly used metal electrodes generated a significant amount of particles during application of pulsed electric field, which could cause precipitation of plasmid DNA from solutions, thereby reducing eTE. For aluminum electrodes, the particles were composed of aluminum hydroxide and/or aluminum oxide, and their median sizes were 300 to 400 nm after the buffer being pulsed 4 to 8 times at 400 V cm, 5 ms duration and 1 Hz frequency. The precipitation could be prevented by using carbon (graphite) electrodes in electrotransfection experiments. The use of carbon electrodes also increased cell viability. Taken together, the study suggested that electrodes made of inner materials were desirable for electrotransfection of cells .
电穿孔转染作为一种通用的非病毒基因递送方法已被广泛应用。然而,与病毒方法相比,电穿孔转染效率(eTE)仍然较低且不稳定。为了了解eTE不稳定的潜在机制,我们研究了电极材料对eTE和哺乳动物细胞活力的影响。该研究数据表明,常用的金属电极在施加脉冲电场期间会产生大量颗粒,这可能导致质粒DNA从溶液中沉淀,从而降低eTE。对于铝电极,颗粒由氢氧化铝和/或氧化铝组成,在缓冲液于400V/cm、5ms持续时间和1Hz频率下脉冲4至8次后,其平均尺寸为300至400nm。在电穿孔转染实验中使用碳(石墨)电极可以防止沉淀。使用碳电极还提高了细胞活力。综上所述,该研究表明内部材料制成的电极对于细胞的电穿孔转染是理想的。