A METHOD FOR THE ISOLATION AND CULTURE OF RAT PERIPHERAL-NERVE VASCULAR ENDOTHELIAL-CELLS

In studying autoimmune diseases of the human peripheral nervous system (PNS), in vitro studies involving the use of cultured rat Schwann cel ls, neurons, and disease-inducing immune system cells have provided ba sic information about disease pathogenesis. For example, T-cells that induce experimental allergic neuritis have been shown in vitro to dama ge Schwann cells, the target cell in these diseases. However, before m aking contact with Schwann cells, these T-cells must first pass throug h the blood-nerve barrier. Despite the importance of this interaction, no studies employing PNS endothelial cells in coculture with dorsal r oot ganglia cells to mimic the environment of the blood-nerve barrier have been reported. This paper describes a simple method for the isola tion and culture of peripheral nerve vascular endothelial cells from a dult rats that should facilitate in vitro studies of the blood-nerve b arrier. Endothelial cells were identified by their expression of an en dothelial cell marker, Factor VIII/von Willebrand factor. Their identi ty was further confirmed by their inability to express Thy 1.1, a fibr oblast marker, and their in vitro morphology. Purity of endothelial ce ll cultures was ensured by a regular program of Thy 1.1 complement dep letion of fibroblasts. (C) 1994 Academic Press, Inc.