Electroporation-mediated delivery of macromolecules to intestinal epithelia

The ability to electroporate cell monolayers could be a powerful tool for studying and treating inflammatory diseases that affect the intestinal epithelium. Electroporation involves the application of short electric field pulses to cells causing the transient disruption of cell membranes and the transport of membrane-impermeant molecules into viable cells. Two human colonic cell lines (Caco-2 and T84) that model the intestinal epithelium were electroporated using calcein and bovine serum albumin (BSA) as marker molecules. Flow cytometry was used to quantify molecular transport and cell viability. Results for both cell types show that there was a general increase in the average uptake of calcein and BSA as the voltage applied and pulse lengths were increased above a quasi-threshold. Cell viability decreased as these two parameters were increased. These trends are consistent with those observed for cell suspensions. In contrast to the vast majority of previous studies which have examined cell suspensions, this study demonstrates that significant uptake of macromolecules can be achieved when electroporating monolayers of intestinal epithelial cells. This is important since monolayers are required to maintain the polarized structure necessary for proper function in culture and represent the correct physiological structure in vivo