Modeling of morphologically realistic large-scale multicellular electroporation for population-level analysis

• A novel automated framework reconstructs realistic large-scale multicellular geometries. • Simulated electroporation in ∼400 HCMEC/D3 cells using a 2-2-2 pulsed electric field protocol. • Revealed electroporation heterogeneity due to cell morphology and field orientation. • Established a scalable pipeline for future realistic 3D electroporation modeling and therapy design. This study explores the dynamics of multicellular electroporation within a monolayer cluster of HCMEC/D3 cells subjected to pulsed electric fields. By proposing an automated approach capable of reconstructing realistic multicellular morphology, we accurately simulate the temporal-spatial distribution of electroporation characteristics, including pore density, pore radius, pore area ratio (PAR), and transmembrane potential (TMP) across the cell and nuclear membranes in large-scale multicellular structures comprising up to ∼400 cells. Our results reveal significant heterogeneity in the electroporation response among cells, primarily driven by variations in cell morphology and orientation relative to the applied electric field. High pore density and PAR are predominantly localized in the polar regions of cells align with the field direction, while regions perpendicular to the electric field exhibit minimal electroporation. Importantly, the results suggests that cell death induced by electroporation is not necessarily contingent on the entire cell membrane permeabilization; rather, it is closely related to the fraction of the membrane affected and the duration of the electroporated state. The findings from this study provide valuable insights into the biophysical mechanisms of electroporation, offering potential pathways for enhancing the precision and effectiveness of electroporation-based therapies. Furthermore, the proposed methodology for reconstructing large-scale 2D multicellular systems lays the groundwork for future extension to realistic 3D tissue models.