Primarily cultured Schwann cells are essential for the investigation of molecular mechanisms regulating proliferation, survival, differentiation, and myelination of Schwann cell and for the development of efficient transplantation for regeneration of injured spinal cord or peripheral nervous system. Here we describe a basic protocol for isolation and purification of primary Schwann cell from neonatal rat or mouse and discuss some modifications adapted to the culturing from adult nerves and optional methods for Schwann cell enrichment.