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Dielectric evolution of mammalian cell membranes after exposure to pulsed electric fields

机译:暴露于脉冲电场后哺乳动物细胞膜的介电演化

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We have investigated the temporal evolution of the dielectric properties of Jurkat cells, a malignant human T-cell line, after exposure to microsecond and nanosecond pulsed electric fields. Suspensions with a 10% volume fraction of Jurkat cells were exposed to either 8 consecutive 1.1 kV/cm electroporation pulses of 100 μs, or 8 consecutive 20-kV/cm pulses of 300 ns, applied with a repetition rate of about 1 Hz. The dielectric spectrum of the cell suspensions before and after exposure was determined by means of time domain reflectometry dielectric spectroscopy. After correction for electrode polarization, the conductivity and permittivity of the cell membrane was derived by fitting the suspension spectrum to a combination of Maxwell-Wagner mixture model and single-shell cell model. Both exposure regimens lead to an increase in the conductivities of suspending medium and cell membrane indicating that membrane poration had occurred. This was confirmed by membrane integrity markers. Significantly higher membrane conductivities without uptake of membrane integrity marker suggest different pore formation mechanisms. Accordingly, dielectric measurements allow us to assess conformational and possibly induced functional changes of cells after exposure on biologically relevant time scales, and in turn, evaluate and compare the efficacy of chosen pulse parameters.
机译:我们已经研究了Jurkat细胞(一种恶性的人类T细胞系)暴露于微秒和纳秒脉冲电场后的介电性能随时间的演变。将Jurkat细胞体积分数为10%的悬浮液暴露于100μs的8个连续的1.1 kV / cm电穿孔脉冲或300 ns的8个连续的20 kV / cm脉冲,重复频率约为1 Hz。借助于时域反射法介电光谱法测定暴露之前和之后细胞悬浮液的介电光谱。校正电极极化后,通过将悬浮谱拟合为Maxwell-Wagner混合模型和单壳细胞模型的组合,得出细胞膜的电导率和介电常数。两种暴露方案均导致悬浮培养基和细胞膜的电导率增加,表明发生了膜渗透。膜完整性标志物证实了这一点。在不吸收膜完整性标记的情况下,较高的膜电导率表明存在不同的孔形成机制。因此,介电测量使我们能够在生物学相关的时间尺度上评估细胞构象化和可能诱导的功能变化,进而评估和比较所选脉冲参数的功效。

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