There is a paucity of information on the roles of extracellular matrix (ECM) and substrate molecules in general with regard to the growth and differentiation of neural stem and progenitor cells. There are well-established findings of a dense, presumably astrocyte-derived ECM in the persistently neurogenic subependymal zone and its migratory extension the rostral migratory stream. Cells cultured from this region, as well as from early postnatal cerebellum, generate multipotent neurospheres, but at present there is little information as to the ECM regulation of these neural stem cell populations. The present study examined the behavior of cerebellar-derived neurospheres on the matrix components laminin, fibronectin, and chondroitin sulfate proteoglycan. The results showed that laminin and fibronectin significantly increase cell migration velocity as compared to CSPG. Fibronectin effected a maximal velocity after 48 h, whereas maximal velocity on laminin and CSPG was not reached until 72 h. Both laminin and fibronectin were very permissive substrates for cellular outgrowth. Chondroitin sulfate proteoglcyan showed a significant inhibition of migratory outgrowth and velocity. These ECM molecules did not appear to affect the fate choice of neurons and glia, thus their role in neuropoietic structures may be to facilitate or deter cell movement and process outgrowth.