Single Cell Forces after Electroporation
Single Cell Forces after Electroporation
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DOI：
10.1021/acsnano.0c07020
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发表时间：
2021-02-23
期刊：
影响因子：
17.1
通讯作者：
Davalos, Rafael, V
中科院分区：
文献类型：
--
作者：
Graybill, Philip M.;Jana, Aniket;Davalos, Rafael, V
外源性高压脉冲通过称为电穿孔的现象增加细胞膜通透性。这个过程也可能破坏细胞骨架，引起细胞收缩性的变化;然而，电穿孔后细胞力的收缩特征仍然未知。在这里，电穿孔后的单细胞力是使用悬浮的细胞外基质模拟纳米纤维作为力传感器来测量的。在三个电压幅度（500、1000和1500 V）和两个方向（平行和垂直于细胞方向）下输送10个100 μ s脉冲，将胶质母细胞瘤细胞暴露于441 V cm（-1）和1366 V cm（-1）之间的电场。电穿孔后细胞膜驱动的力损失和恢复涉及三个不同的阶段。低电场强度不会引起破坏，但较高的电场几乎消除了电穿孔后2-10分钟的收缩性，因为细胞在钙介导的收缩后变圆（阶段1）。在圆整后，大多数分析的细胞进入不寻常的和意想不到的双相阶段（阶段2），其特征在于电穿孔后数十分钟的收缩性增加，随后是力松弛。双相阶段与肌动蛋白破坏驱动的水泡同时发生。最后，细胞伸长并在1-3小时内恢复其电穿孔前的形态和收缩性（阶段3）。随着垂直于细胞方向施加的电压的增加，我们观察到细胞活力的显著下降。多个健康和癌细胞系的实验表明，收缩力是比细胞形状对电穿孔更动态和敏感的度量。对电穿孔后细胞收缩性的机械生物学理解将加深我们对驱动恢复的机制的理解，并可能对分子医学，遗传工程和细胞生物物理学产生影响。
Exogenous high-voltage pulses increase cell membrane permeability through a phenomenon known as electroporation. This process may also disrupt the cell cytoskeleton causing changes in cell contractility; however, the contractile signature of cell force after electroporation remains unknown. Here, single-cell forces post-electroporation are measured using suspended extracellular matrix-mimicking nanofibers that act as force sensors. Ten, 100 mu s pulses are delivered at three voltage magnitudes (500, 1000, and 1500 V) and two directions (parallel and perpendicular to cell orientation), exposing glioblastoma cells to electric fields between 441 V cm(-1) and 1366 V cm(-1). Cytoskeletal-driven force loss and recovery post-electroporation involves three distinct stages. Low electric field magnitudes do not cause disruption, but higher fields nearly eliminate contractility 2-10 min post-electroporation as cells round following calcium-mediated retraction (stage 1). Following rounding, a majority of analyzed cells enter an unusual and unexpected biphasic stage (stage 2) characterized by increased contractility tens of minutes post-electroporation, followed by force relaxation. The biphasic stage is concurrent with actin disruption-driven blebbing. Finally, cells elongate and regain their pre-electroporation morphology and contractility in 1-3 h (stage 3). With increasing voltages applied perpendicular to cell orientation, we observe a significant drop in cell viability. Experiments with multiple healthy and cancerous cell lines demonstrate that contractile force is a more dynamic and sensitive metric than cell shape to electroporation. A mechanobiological understanding of cell contractility post-electroporation will deepen our understanding of the mechanisms that drive recovery and may have implications for molecular medicine, genetic engineering, and cellular biophysics.