Subcultures have been established from primary rat brain cell cultures and have been characterised with a range of cell-specific immunocytochemical markers. The subcultures are mainly composed of fibrous astrocytes, oligodendrocytes and neurones. The cells do not divide to any great extent giving a system where it is possible to follow culture development at the cellular level for a number of weeks. During this time oligodendrocytes colonise subpopulations of neurones, differentiate further showing the presence of myelin basic protein and elaborate myelin-like membrane; the fibrous astrocytes remain scattered uniformly throughout the cultures. Radially oriented processes emerge from the oligodendrocyte-neurone aggregates which subsequently coalesce to form fascicles that link the clusters of cells together. These fascicles react with antibodies for both neurofilament protein and myelin basic protein. The subcultures provide a straightforward system that is composed of cells derived entirely from the CNS, is free from mitotic inhibitors and yet retains a sufficiently low cell density to allow immunocytochemical identification of the cell types present. The subcultures should be useful for the study of trophic interactions between oligodendrocytes and neurones as well as the early events associated with myelinogenesis.