The role of lipoproteins secreted by cortical glial cells in axon growth of central nervous system (CNS) neurons was investigated. We first established compartmented cultures of CNS neurons (retinal ganglion cells). Addition of glial cell-conditioned medium (GCM) to distal axons increased the rate of axon extension by approximately 50%. Inhibition of 3-hydroxy-3-methylglutaryl-CoA reductase in glial cells diminished the secretion of cholesterol and apolipoprotein E, and prevented the growth stimulatory effect of GCM. When glia-derived lipoproteins containing apolipoprotein E were provided to distal axons, axon extension was stimulated to the same extent as by GCM. In contrast, addition of lipoproteins to cell bodies failed to enhance growth. The growth stimulatory effect of glial lipoproteins was abrogated in the presence of receptor-associated protein, RAP, indicating involvement of receptor(s) of the low density lipoprotein receptor family in stimulation of axonal extension. These observations suggest that glial cells stimulate axon growth of CNS neurons by providing lipoproteins containing cholesterol and apolipoprotein E to distal axons.