Abstract：
OBJECTIVE: During physical transfection, an electrical field or mechanical force is used to induce cell transfection. We tested if the disruption of a dense actin layer underneath the membrane of a suspended cell enhances cell transfection.
RESULTS: A bubble generator was used to electromechanically stimulate suspended cells. To clarify the influence of the actin layer (the actin cortex) on cell transfection efficiency, we used an actin polymerization inhibitor (cytochalasin D) to disrupt the actin cortex before electromechanical stimulation. Without cytochalasin D treatment, signals from the overall actin cortex decreased after electromechanical stimulation. With cytochalasin D treatment, there was localized F-actin aggregation under static conditions. After electromechanical stimulation, there was a partial loss (localized disruption), but no overall disruption, of the actin cortex. With the pretreatment with cytochalasin D, the transfection efficiency of plasmids (4.7, 8.3, or 11 kbp) into NIH/3T3 or UMR-106 cells increased significantly after exposure to electromechanical stimulation.
CONCLUSIONS: Localized distribution of the actin cortex before exposure to electromechanical stimulation is crucial for inducing a partial loss of the cortex, which improves transfection efficiency and large plasmid delivery.
RESULTS: A bubble generator was used to electromechanically stimulate suspended cells. To clarify the influence of the actin layer (the actin cortex) on cell transfection efficiency, we used an actin polymerization inhibitor (cytochalasin D) to disrupt the actin cortex before electromechanical stimulation. Without cytochalasin D treatment, signals from the overall actin cortex decreased after electromechanical stimulation. With cytochalasin D treatment, there was localized F-actin aggregation under static conditions. After electromechanical stimulation, there was a partial loss (localized disruption), but no overall disruption, of the actin cortex. With the pretreatment with cytochalasin D, the transfection efficiency of plasmids (4.7, 8.3, or 11 kbp) into NIH/3T3 or UMR-106 cells increased significantly after exposure to electromechanical stimulation.
CONCLUSIONS: Localized distribution of the actin cortex before exposure to electromechanical stimulation is crucial for inducing a partial loss of the cortex, which improves transfection efficiency and large plasmid delivery.
摘要：
目的：在物理转染过程中，电场或机械力用于诱导细胞转染。我们测试了悬浮细胞膜下方致密肌动蛋白层的破坏是否增强了细胞转染。
结果:使用气泡发生器机电刺激悬浮细胞。为了阐明肌动蛋白层（肌动蛋白皮层）对细胞转染效率的影响，我们使用肌动蛋白聚合抑制剂（细胞松弛素D）在电机械刺激之前破坏肌动蛋白皮质。没有细胞松弛素D治疗，机电刺激后，来自整个肌动蛋白皮质的信号降低。用细胞松弛素D治疗，在静态条件下存在局部F-肌动蛋白聚集。机电刺激后，有部分损失(局部中断)，但没有整体中断，肌动蛋白皮质。用细胞松弛素D预处理，暴露于机电刺激后，质粒（4.7、8.3或11kbp）转染NIH/3T3或UMR-106细胞的效率显着提高。
结论：肌动蛋白皮质在暴露于机电刺激之前的局部分布对于诱导皮质的部分损失至关重要，这提高了转染效率和大质粒递送。
结果:使用气泡发生器机电刺激悬浮细胞。为了阐明肌动蛋白层（肌动蛋白皮层）对细胞转染效率的影响，我们使用肌动蛋白聚合抑制剂（细胞松弛素D）在电机械刺激之前破坏肌动蛋白皮质。没有细胞松弛素D治疗，机电刺激后，来自整个肌动蛋白皮质的信号降低。用细胞松弛素D治疗，在静态条件下存在局部F-肌动蛋白聚集。机电刺激后，有部分损失(局部中断)，但没有整体中断，肌动蛋白皮质。用细胞松弛素D预处理，暴露于机电刺激后，质粒（4.7、8.3或11kbp）转染NIH/3T3或UMR-106细胞的效率显着提高。
结论：肌动蛋白皮质在暴露于机电刺激之前的局部分布对于诱导皮质的部分损失至关重要，这提高了转染效率和大质粒递送。
