Glial-conditioned medium and attachment to ConA are essential for long-term culture of cortical neurons

Cortical brain cells from 14-day-old mouse embryos were seeded on various substrates and cultivated in serum-free medium with or without conditioned medium from astrocytes or C6 glioma cells. Poly-L-lysine was shown to be the best substrate for cell attachment followed by Concanavalin A (ConA) and adhesion particles derived from glia cells. Cells grown on ConA sprouted rapidly and formed large networks. Survival of neurons was greatly prolonged when glia-conditioned medium (GCM) was present in the culture medium. Cells grown on ConA were then viable for more than 4 weeks. Without GCM, neurons survived in culture for about 2 weeks, regardless of the substrate. Endothelial cell growth supplement or acidic fibroblast growth factor increased survival of neurons but also stimulated proliferation of astrocytes.