Isolation of rat Schwann cells based on cell sorting.

Abstract

The present study presented a protocol that can be used to obtain rapidly a high purity of proliferating rat Schwann cells from freshly dissociated rat peripheral nerves. The sciatic nerves of newborn rats (1–3 day old) were dissociated, and the Schwann cells (SCs) were purified using fluorescence-activated cell sorting (FACS) based on the SC membrane-specific expression of the low-affinity nerve growth factor receptor, p75NGFR and oligodendrocyte marker 4. Following sorting, the cells were plated on poly-l-lysine-coated dishes in SC culture medium containing DMEM with 10% FBS, 1% penicillin/streptomycin, 2 µM forskolin and 10 ng/ml HRG. The purified rat SCs were propagated for passaging until confluent. This protocol resulted in SC cultures, which were >98% pure. This FACS-based protocol can be used to facilitate future investigations of general SC biology.