2024 AIChE Annual Meeting
(175az) Elucidating the Geometric and Charge Transfer Effects on the Spatial Uniformity of the Nanopore Electroporation Cell Transfection Method
Transfection, the process of delivering macromolecules such as DNA and mRNA into cells, has enabled researchers to engineer cells for a variety of medical applications including cellular manipulation, cellular imaging, and genome engineering. Nanopore electroporation (NanoEP) was developed recently as a gentle and highly efficient transfection alternative to bulk electroporation (EP). In bulk EP, an applied electric field causes pores to form throughout the cell membrane, but can cause high cell death rate due to the high voltage felt along the entirety of the cell membrane. NanoEP, on the other hand, utilizes an insulating nanoporous polycarbonate track-etched membrane that localizes the electric field only along the cell membrane over the nanopores, significantly improving delivery efficiency and cell viability. Cell coverage and pore distribution of the nanoporous insulated membrane are known factors that can affect the overall uniformity of delivery in NanoEP. In this study, we investigated additional factors such as device geometry, electrode proximity, and electrode charge transfer resistance in NanoEP systems to enhance the cell delivery/depletion uniformity using bioactive organic molecules and proteins such as calcein and bovine serum albumin. These new design criteria further establish NanoEP as a facile and scalable transfection method with high uniformity, efficiency, and viability for various biomedical applications.
