Department of Electronics, University of York, Heslington, York YO10 5DD, UK.
Phys Med Biol. 2010 Feb 21;55(4):1219-29. doi: 10.1088/0031-9155/55/4/021. Epub 2010 Feb 2.
This paper describes a method of effectively electroporating mammalian cell membranes with pulsed alternating-current (ac) electric fields at field strengths of 30-160 kV m(-1). Although many in vivo electroporation protocols entail applying square wave or monotonically decreasing pulses via needles or electrode plates, relatively few have explored the use of pulsed ac fields. Following our previous study, which established the effectiveness of ac fields for electroporating cell membranes, a primary/secondary coil system was constructed to produce sufficiently strong electric fields by electromagnetic induction. The primary coil was formed from the applicator of an established transcranial magnetic stimulation (TMS) system, while the secondary coil was a purpose-built device of a design which could eventually be implanted into tissue. The effects of field strength, pulse interval and cumulative exposure time were investigated using microscopy and flow cytometry. Results from experiments on concentrated cell suspensions showed an optimized electroporation efficiency of around 50%, demonstrating that electroporation can be practicably achieved by inducing such pulsed ac fields. This finding confirms the possibility of a wide range of in vivo applications based on magnetically coupled ac electroporation.
本文描述了一种在 30-160kV/m 的场强下,用脉冲交流(ac)电场有效电穿孔哺乳动物细胞膜的方法。尽管许多体内电穿孔方案都需要通过针或电极板施加方波或单调递减的脉冲,但很少有研究探索使用脉冲 ac 场。在我们之前的研究确定了 ac 场对细胞膜电穿孔的有效性之后,构建了一个初级/次级线圈系统,通过电磁感应产生足够强的电场。初级线圈由已建立的经颅磁刺激(TMS)系统的应用器形成,而次级线圈是一个专门设计的设备,其设计最终可以植入组织中。通过显微镜和流式细胞术研究了场强、脉冲间隔和累积暴露时间的影响。在浓缩细胞悬浮液上的实验结果表明,优化的电穿孔效率约为 50%,表明通过感应这种脉冲 ac 场可以实际实现电穿孔。这一发现证实了基于磁耦合 ac 电穿孔的广泛体内应用的可能性。