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Most studies of peripheral nerve myelination using culture models are performed with dorsal root ganglion neurons and Schwann cells pre‐purified from the rat. The potential of this model is severely compromised by the lack of rat myelin mutants and the published protocols work poorly with mouse cells, for which numerous myelin mutants are available. This is partly due to difficulties in obtaining sufficient quantities of myelination‐competent mouse Schwann cells. Here, we describe the isolation, purification and expansion of wild‐type, myelination‐competent Schwann cells from the sciatic nerves of 4‐day‐old mouse pups. The method consistently yields 1.9–3.3 × 10⁶ of ∼95% pure Schwann cells from the sciatic nerves of 12–15 4‐day‐old mouse pups, within 14–20 days. The Schwann cell proliferation rate ranges from 2.7‐ to 4.30‐fold growth/week. Proliferation ceases within 4 weeks, when the cells …