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不同活力检测方法揭示不同电穿孔脉冲参数导致的细胞死亡动力学。

Dynamics of Cell Death Due to Electroporation Using Different Pulse Parameters as Revealed by Different Viability Assays.

机构信息

The State Key Laboratory of Power Transmission Equipment and System Security and New Technology, School of Electrical Engineering, Chongqing University, Chongqing, 400044, China.

Faculty of Electrical Engineering, University of Ljubljana, Tržaška 25, 1000, Ljubljana, Slovenia.

出版信息

Ann Biomed Eng. 2024 Jan;52(1):22-35. doi: 10.1007/s10439-023-03309-8. Epub 2023 Sep 13.

Abstract

The mechanisms of cell death due to electroporation are still not well understood. Recent studies suggest that cell death due to electroporation is not an immediate all-or-nothing response but rather a dynamic process that occurs over a prolonged period of time. To investigate whether the dynamics of cell death depends on the pulse parameters or cell lines, we exposed different cell lines to different pulses [monopolar millisecond, microsecond, nanosecond, and high-frequency bipolar (HFIRE)] and then assessed viability at different times using different viability assays. The dynamics of cell death was observed by changes in metabolic activity and membrane integrity. In addition, regardless of pulse or cell line, the dynamics of cell death was observed only at high electroporation intensities, i.e., high pulse amplitudes and/or pulse number. Considering the dynamics of cell death, the clonogenic assay should remain the preferred viability assay for assessing viability after electroporation.

摘要

由于电穿孔导致的细胞死亡的机制仍未被很好地理解。最近的研究表明,电穿孔导致的细胞死亡不是一种即时的全有或全无的反应,而是一个在较长时间内发生的动态过程。为了研究细胞死亡的动力学是否取决于脉冲参数或细胞系,我们用不同的脉冲(单极毫秒、微秒、纳秒和高频双极(HFIRE))处理不同的细胞系,并在不同的时间使用不同的活力测定法评估其活力。通过代谢活性和细胞膜完整性的变化来观察细胞死亡的动力学。此外,无论脉冲或细胞系如何,只有在高电穿孔强度下才会观察到细胞死亡的动力学,即高脉冲幅度和/或脉冲数。考虑到细胞死亡的动力学,克隆形成测定法应该仍然是评估电穿孔后细胞活力的首选活力测定法。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/093f/10761553/ab428f3f53f7/10439_2023_3309_Fig1_HTML.jpg

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