Characterization of neurite outgrowth on electrically stimulated fibroblasts and endothelial cells

Previous work has shown that neurons respond to the presence of an exogenous electric field with increased neurite outgrowth. In addition, in the presence of electrically stimulated Schwann cells, neurite outgrowth also increases. The present study examines the characteristics of neurite response on electrically stimulated monolayers of other cells that would be present in the surrounding in vivo environment following peripheral nerve injury, including fibroblasts (FB) and endothelial cells (EC). FBs and ECs are seeded in sub-confluent monolayers and then subjected to a constant current applied DC electric field (EF) for 8 hours at 50mV/mm. Cells were then allowed to recover in new media for 12 hours. Following recovery primary dissociated neurons were seeded on EC and FB monolayers for 12 hours and then fixed, immunostained, and imaged using fluorescent microscopy. Initial visual observations of neurite extension on stimulated EC monolayers indicates that there may be a slight increase in neurite outgrowth on EF stimulated cells. Currently trials are underway to increase sample size for statistical significance and morphometric data of neurite outgrowth.