Elsevier

Available online 13 May 2020, 107550

Bioelectrochemistry

Highlights:

Cell death occurring following electroporation can be subdivided in two stages.

Membrane resealing or permanent loss of barrier function occurs within 35 minutes.

Cell electroporation leads to loss of variety of intracellular compounds.

Cell viability following electroporation can be increased by adding intracellular compounds.

Abstract

Control of membrane permeability to exogenous compounds by membrane electroporation can lead to cell death, which is related to permanent membrane damage, oxidation stress, leakage of intracellular molecules. In this study, we show that the predominant cell death modality after the application of high voltage electric pulses is related with inability to reseal of initial pores (first stage irreversible electroporation, FirEP). After moderately strong electric pulses, initial pores reseal, however, some cell still die later on due to electric field induced cell stress which leads to delayed cell death (late-stage irreversible electroporation, LirEP). According to our data, the period in which the majority of cells commit to either pore resealing or complete loss of barrier function depends on the intensity of electric field treatment but did not exceed 35 min. Additionally, we show that after electroporation using electric pulse parameters that induce LirEP, some cells can be rescued by supplementing medium with compounds obtained from irreversibly electroporated cells. We determined that the intracellular molecules that contribute to the increase of cell viability are larger than 30 kDa. This serves to prove that the loss of intracellular compounds plays a significant role in the decrease of cell viability after electroporation.

Keywords

irreversible electroporation

cell death

calcein

ATP

propidium iodide

pore resealing

supernatant

intracellular molecules

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