A method for obtaining Schwann cell cultures from adult rabbit nerve based on “in vitro” pre-degeneration and neuregulin treatment

Abstract

Schwann cells (SCs) are basic elements for cell therapy and tissue engineering in the central and peripheral nervous system. Therefore, the development of a reliable method to obtain SC cultures is required. For possible therapeutic applications the cultures need to produce a sufficiently large number of SCs with a high level of purity in a relatively short period of time. To increase SC yield and purity we pre-degenerated pieces of 1-2 mm of adult rabbit sciatic nerves by incubating them for seven days in Dulbecco’s Modified Eagle’s Medium supplemented with 10% fetal bovine serum, penicillin/streptomycin and NRG1-ß1. Following pre-degeneration the nerve pieces were dissociated and then cultured for 6 or 15 days in the same culture medium. After 6 days of culture we obtained around 9.5x103 cells/mg with approximately 94% SCs (S-100 positive) purity. After 15 days of culture the yield was about 80x103 cells/mg and the purity was approximately 75%. Pre-degeneration and subsequent culture of small pieces of adult nerve with NRG1-ß1 supplemented medium increased the number of SCs and restricted the overgrowth of fibroblast-like cells