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

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Abstract
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.