Improvement in Electrotransfection of Cells Using Carbon-Based Electrodes.
Improvement in Electrotransfection of Cells Using Carbon-Based Electrodes.
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DOI：
10.1007/s12195-016-0452-9
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发表时间：
2016-12
影响因子：
2.8
通讯作者：
Yuan F
中科院分区：
文献类型：
--
作者：
Chang CC;Mao M;Liu Y;Wu M;Vo-Dinh T;Yuan F
电转染作为一种通用的、非病毒的基因传递方法已被广泛使用。然而，与病毒方法相比，电转染效率（eTE）仍然较低且不稳定。为了了解不稳定电激反应的潜在机制，我们研究了电极材料对电激反应和哺乳动物细胞活力的影响。研究数据表明，在脉冲电场作用下，常用的金属电极会产生大量颗粒，导致质粒DNA从溶液中析出，从而降低eTE。对于铝电极，缓冲液在400 V cm - 1,5 ms脉冲4 ~ 8次后，其中位尺寸为300 ~ 400 nm，由氢氧化铝和/或氧化铝组成。在电转染实验中使用碳（石墨）电极可以防止沉淀。碳电极的使用也提高了细胞活力。综上所述，该研究表明，由内部材料制成的电极是体外细胞电转染的理想选择。
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−1, 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 in vitro.