title: Water influx and cell swelling after nanosecond electropermeabilization.
abstract: Pulsed electric fields are used to permeabilize cell membranes in biotechnology and the clinic. Although molecular and continuum models provide compelling representations of the mechanisms underlying this phenomenon, a clear structural link between the biomolecular transformations displayed in molecular dynamics (MD) simulations and the micro- and macroscale cellular responses observed in the laboratory has not been established. In this paper, plasma membrane electropermeabilization is characterized by exposing Jurkat T lymphoblasts to pulsed electric fields less than 10ns long (including single pulse exposures), and by monitoring the resulting osmotically driven cell swelling as a function of pulse number and pulse repetition rate. In this way, we reduce the complexity of the experimental system and lay a foundation for gauging the correspondence between measured and simulated values for water and ion transport through electropermeabilized membranes. We find that a single 10MV/m pulse of 5ns duration produces measurable swelling of Jurkat T lymphoblasts in growth medium, and we estimate from the swelling kinetics the ion and water flux that follows the electropermeabilization of the membrane. From these observations we set boundaries on the net conductance of the permeabilized membrane, and we show how this is consistent with model predictions for the conductance and areal density of nanoelectropulse-induced lipid nanopores.
authors.affiliation: CNR-Institute for Electromagnetic Sensing of Environment, Naples, Italy. romeo.s@irea.cnr.it
authors.forename: Stefania
authors.initials: S
authors.lastname: Romeo
authors.forename: Yu-Hsuan
authors.initials: YH
authors.lastname: Wu
authors.forename: Zachary A
authors.initials: ZA
authors.lastname: Levine
authors.forename: Martin A
authors.initials: MA
authors.lastname: Gundersen
authors.forename: P Thomas
authors.initials: PT
authors.lastname: Vernier
mesh_terms: D049108:Cell Enlargement; D002462:Cell Membrane / Q000378:metabolism*; D002463:Cell Membrane Permeability; D018274:Electroporation; D006801:Humans; D019169:Jurkat Cells; D007700:Kinetics; D008051:Lipid Bilayers; D008856:Microscopy, Fluorescence; D056004:Molecular Dynamics Simulation; D036103:Nanotechnology; D014867:Water / Q000378:metabolism*
chemical_list: D008051:Lipid Bilayers; D014867:Water