Magnetic cell sorting for enriching Schwann cells from adult mouse peripheral nerves.
We have devised a simple method to purify mitotically active Schwann 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.
Manent
Jan
J
INSERM U434, Fondation Jean Dausset-CEPH, 27, rue Juliette Dodu, 75010 Paris, France.
Oguievetskaia
Ksenia
K
Bayer
Jan
J
Ratner
Nancy
N
Giovannini
Marco
M
D000818:Animals; D002455:Cell Division / Q000502:physiology; D002478:Cells, Cultured; D005434:Flow Cytometry / Q000379:methods*; D008280:Magnetics / Q000295:instrumentation*; D051379:Mice; D010525:Peripheral Nerves / Q000166:cytology* / Q000378:metabolism; D012583:Schwann Cells / Q000166:cytology* / Q000378:metabolism