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Ionomycin-Induced Changes in Membrane Potential Alter Electroporation Outcomes in HL-60 Cells

机译:碘霉素诱导的HL-60细胞膜电位改变改变电穿孔的结果。

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摘要

Previous studies have shown greater fluorophore uptake during electroporation on the anode-facing side of the cell than on the cathode-facing side. Based on these observations, we hypothesized that hyperpolarizing a cell before electroporation would decrease the requisite pulsed electric field intensity for electroporation outcomes, thereby yielding a higher probability of reversible electroporation at lower electric field strengths and a higher probability of irreversible electroporation (IRE) at higher electric field strengths. In this study, we tested this hypothesis by hyperpolarizing HL-60 cells using ionomycin before electroporation. These cells were then electroporated in a solution containing propidium iodide, a membrane integrity indicator. After 20 min, we added trypan blue to identify IRE cells. Our results showed that hyperpolarizing cells before electroporation alters the pulsed electric field intensity thresholds for reversible electroporation and IRE, allowing for greater control and selectivity of electroporation outcomes.
机译:先前的研究表明,在电穿孔过程中,细胞在阳极侧的吸收的荧光团要多于阴极侧。基于这些观察,我们假设在电穿孔之前对细胞进行超极化将降低电穿孔结果所需的脉冲电场强度,从而在较低的电场强度下产生较高的可逆电穿孔可能性,而在较高的电场强度下产生较高的不可逆电穿孔(IRE)可能性。电场强度。在这项研究中,我们通过在电穿孔前使用离子霉素对HL-60细胞进行超极化来检验该假设。然后将这些细胞在含有碘化丙锭(膜完整性指示剂)的溶液中进行电穿孔。 20分钟后,我们加入了台盼蓝以识别IRE细胞。我们的结果表明,电穿孔前的超极化细胞会改变可逆电穿孔和IRE的脉冲电场强度阈值,从而实现对电穿孔结果的更大控制和选择性。

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