High-Throughput Capable Three-Dimensional Tissue Model for Quantification of Electroporation Thresholds

Electroporation is a promising technology utilizing electrical pulses for macromolecule delivery and soft-tissue ablation, with applications that include next-generation prophylactics and the treatment of genetic diseases such as cancer. This study demonstrates a high-throughput capable 3D tissue culture model for quantification of the reversible and irreversible electroporation thresholds for a given electroporation protocol. By using a non-uniform electric field and analyzing the spatial distribution of transfected cells, both reversible and irreversible thresholds can be identified within a single sample, increasing the efficiency at which electroporation protocols can be characterized, especially for in vivo translation. To show this capability, 3D tissue mimics containing HEK293 cells were transfected using a ring and pin electrode to deliver a GFP-encoding plasmid. Electroporation thresholds were then derived based on fluorescent microscopy images of the transfected samples. This model demonstrates potential for use as a means for high-throughput evaluation of electroporation protocols, a key advantage over current methods to evaluate these thresholds, which tend to be time-intensive and are less representative of in vivo conditions.