The meninges (the pia mater and the arachnoid) covering the surface of brain parenchyma are known to participate in the regulation of blood circulation and the blood-cerebrospinal fluid barrier. In the present study, we isolated and characterized some of the major proteins secreted into the conditioned medium of the meningeal cell cultures prepared from 1-2-day-old rats. Five protein molecules (7 kDa, 16 kDa, 18 kDa, 27 kDa and 32 kDa) were recognized as major proteins in the conditioned medium by SDS-PAGE. The major proteins were isolated and purified to homogeneity, respectively. Their N-terminal sequences, except that of 27 kDa protein, were successfully determined. Homology search has revealed that the N-terminal sequences of the 7 kDa protein and the 32 kDa protein were identical with those of insulin-like growth factor-II (IGF-II) and IGF-binding protein-2 (IGFBP-2), respectively. The N-terminal sequences of the 16 kDa and 18 kDa molecules were identical with those of the corresponding fragments of IGFBP-2. The present study demonstrates that cultured meningeal cells produce and secrete large amounts of IGF-II and IGFBP-2. The meninges may, therefore, be one of the main sources of these proteins in the cerebrospinal fluid and involved in the modulation of neuronal and/or glial cell survival or functioning.