University of Ljubljana, Faculty of Electrical Engineering, Tržaška 25, 1000 Ljubljana, Slovenia.
University of Ljubljana, Faculty of Electrical Engineering, Tržaška 25, 1000 Ljubljana, Slovenia.
Bioelectrochemistry. 2019 Dec;130:107342. doi: 10.1016/j.bioelechem.2019.107342. Epub 2019 Aug 1.
Electroporation is a method which uses an adequate number of electric pulses of enough amplitude, duration and number applied to cells, thus inducing transient permeabilization of the cell membrane. Due to possibility that microenvironment in applications of in vivo electroporation is slightly acidic, we studied the effects of slightly acidic electroporation and recovery medium. We observed no difference in the permeabilization threshold, detected by propidium iodide, of cells which were electroporated and allowed to recover in growth (pH 7.8) or acidic (pH 6.5) medium. In contrast, statistically significant difference was observed in survival of cells that were exposed to pulse amplitudes greater than permeabilization threshold. Survival of cells was greater if acidic electroporation and recovery medium were used, but acidic extracellular pH decreased gene electrotransfer efficiency. We also observed differences in morphology between cells that were electroporated and left to recover in growth medium and cells that were electroporated and left to recover in acidic medium. Our results imply that slightly acidic extracellular pH allows more efficient repair of damage that is induced on cell membrane during electroporation with high pulse amplitudes.
电穿孔是一种方法，它使用足够数量、足够幅度、足够时长和足够数量的电脉冲施加于细胞，从而诱导细胞膜瞬时通透性。由于在体内电穿孔应用中微环境可能略微呈酸性，我们研究了略微酸性的电穿孔和恢复介质的影响。我们观察到，在电穿孔并在生长（pH 7.8）或酸性（pH 6.5）介质中恢复的细胞中，碘化丙啶检测到的通透性阈值没有差异。相比之下，在暴露于大于通透性阈值的脉冲幅度的细胞的存活方面观察到了统计学上显著的差异。如果使用酸性电穿孔和恢复介质，则细胞的存活率更高，但细胞外 pH 值降低会降低基因电转移效率。我们还观察到，在电穿孔并在生长培养基中恢复的细胞与电穿孔并在酸性培养基中恢复的细胞之间的形态存在差异。我们的结果表明，细胞外略微酸性的 pH 值允许在高脉冲幅度下进行电穿孔时对细胞膜诱导的损伤进行更有效的修复。
