A Combined Numerical and Experimental Investigation of Localized Electroporation-based Transfection and Sampling

Abstract

Localized electroporation has evolved as an effective technology for the delivery of foreign molecules into adherent cells, and more recently, for the sampling of cytosolic content from a small population of cells. Unlike bulk electroporation, where the electric field is poorly controlled, localized electroporation benefits from the spatial localization of the electric field on a small areal fraction of the cell membrane, resulting in efficient molecular transport and high cell-viability.Although there have been numerous experimental reports, a mechanistic understanding of the different parameters involved in localized electroporation is lacking. In this work, we developed a multiphysics model that a) predicts the electro-pore distribution in response to the local transmembrane potential and b) calculates the molecular transport into and out of the cell based on the predicted pore-sizes. Using the model, we identify that cell membrane tension plays a crucial role in enhancing both the amount and the uniformity of molecular transport, particularly for large proteins and plasmids. We qualitatively validate the model predictions by delivering large molecules (fluorescent-tagged bovine serum albumin and mCherry encoding plasmid) and by sampling an exogeneous protein (tdTomato) in an engineered cell line. The findings presented here should inform the future design of microfluidic devices for localized electroporation based sampling, eventually paving the way for temporal, single-cell analysis.

Intracellular delivery of therapeutic proteins. New advancements and future directions

“…Most of the physical approaches for overcoming cell membrane deal with chemical (Stewart et al, 2018) membrane disruption (Mukherjee et al, 2018) or perforation (Chen N. et al, 2022). Although membrane perforation with electroporation (Mukherjee et al, 2018) and microinjection (Keppeke et al, 2015;Chen N. et al, 2022) or sonoporation (Togtema et al, 2012) is the most straightforward method for cytosolic delivery, these strategies are highly efficient in in vitro studies (Tan et al, 2022), but generally toxic, only suitable for introducing a small number of specific proteins into incubated cells and can hardly be used in vivo.…”