Magnetic cell sorting for enriching Schwann cells from adult mouse peripheral nerves

We have devised a simple method to purify mitotically active Schwarm cells (SC) from peripheral nerves of adult mice. Nerves were predegenerated in vitro for 7 days and after dissociation cells were plated on poly-L-lysine/laminin coated dishes in N2 serum-free culture medium supplemented with forskolin and heregulin-beta1. Primary cultures were purified from contaminating fibroblasts by magnetic cell sorting (MACS) based on SC membrane specific expression of p75(NGFR) and enriched to about 99% of SC after MACS from 34 to 91% before sorting. After sorting, purified adult mouse SC were propagated for three passages until confluent to a total surface of 160 cm(2) per mouse (two sciatic and two trigeminal nerves). In addition, we show that this method can be used to purify tumoral SC from mouse NF2-related schwannomas. (C) 2002 Elsevier Science B.V. All rights reserved.