Cells are isolated from sciatic nerve explants of mice or rats. Adherent cultures express Schwann cell-specific markers and have low fibroblast contamination.
Sciatic nerves are removed from the mice and dissected, with fine forceps, to remove the epi- and perineurium. The dissected nerve bundles are then cut into small pieces which are then plated on poly-d-lysine coated plates. Cells are then cultured in a minimal amount of medium consisting of DMEM/F12 supplemented with fetal calf serum (FCS, 10%), penicillin-streptomycin, and L-glutamine, for 2 hours to allow attachment of of the nerve fibers. After attachment of the nerve fibers to the plate, more culture medium is added and cells are cultured for 14 days. Medium is replaced every 2 to 3 days.