<br>基于局部电穿孔的细胞转染和采样的数值与实验相结合的研究,ACS Nano

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基于局部电穿孔的细胞转染和采样的数值与实验相结合的研究
ACS Nano ( IF 15.8 ) Pub Date : 2018-11-19 00:00:00 , DOI: 10.1021/acsnano.8b05473
Prithvijit Mukherjee , S Shiva P Nathamgari , John A Kessler ₁ , Horacio D Espinosa

Affiliation

局部电穿孔已发展成为一种有效的技术，可将外源分子输送到细胞中，同时保持其活力。因此，该技术在活细胞内容物采样和单细胞水平上细胞状态的时间评估方面具有潜在的应用。尽管已有大量关于基于局部电穿孔的递送的实验报告，但缺乏对该过程的机械理解阻碍了其在采样中的实施。在这项工作中，我们开发了一个多物理场模型，可以预测局部电穿孔期间分子进出细胞的运输。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、细胞汇合度和基底中纳米通道的密度，以便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起着至关重要的作用，特别是对于大分子而言。我们通过提供大分子（牛血清白蛋白和 mCherry 编码质粒）并通过在工程细胞系中采样外源蛋白 (tdTomato) 来定性验证局部电穿孔平台上的模型预测。

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

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.

更新日期：2018-11-19

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