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Combined Numerical and Experimental Investigation of Localized Electroporation-Based Cell Transfection and Sampling

机译：基于电穿孔的局部细胞转染和采样的组合数值和实验研究

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摘要

Localized electroporation has evolved as an effective technology for the delivery of foreign molecules into cells while preserving their viability. Consequently, this technique has potential applications in sampling the contents of live cells and the temporal assessment of cellular states at the single-cell level. Although there have been numerous experimental reports on localized electroporation-based delivery, a lack of a mechanistic understanding of the process hinders its implementation in sampling. In this work, we develop a multiphysics model that predicts the transport of molecules into and out of the cell during localized electroporation. Based on the model predictions, we optimize experimental parameters such as buffer conditions, electric field strength, cell confluency, and density of nanochannels in the substrate for successful delivery and sampling via localized electroporation. We also identify that cell membrane tension plays a crucial role in enhancing both the amount and the uniformity of molecular transport, particularly for macromolecules. We qualitatively validate the model predictions on a localized electroporation platform by delivering large molecules (bovine serum albumin and mCherry-encoding plasmid) and by sampling an exogeneous protein (tdTomato) in an engineered cell line.
机译：局部电穿孔已发展成为一种有效的技术，可在保持外来分子活力的同时将其输送到细胞中。因此，该技术在采样活细胞的内容以及单细胞水平的细胞状态的时间评估方面具有潜在的应用。尽管有许多关于基于电穿孔的局部递送的实验报告，但是对该过程缺乏机械理解会阻碍其在采样中的实施。在这项工作中，我们开发了一个多物理场模型，该模型可以预测局部电穿孔过程中分子进出细胞的转运。基于模型预测，我们优化了实验参数，例如缓冲条件，电场强度，细胞融合度和底物中纳米通道的密度，以通过局部电穿孔成功递送和采样。我们还发现，细胞膜张力在增强分子运输的数量和均匀性方面都起着至关重要的作用，特别是对于大分子而言。我们通过传递大分子（牛血清白蛋白和mCherry编码质粒）并在工程细胞系中取样外源蛋白质（tdTomato），在局部电穿孔平台上定性验证模型预测。

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