1. Morphologically developmental properties of fetal mouse cortical neurons in the chemically defined serum- and antioxidant-free culture condition were observed. Also, cellular composition in cultures was identified by immunostaining with anti-NSE and anti-GFAP.
2. Various cell densities ranging from 1 × 103 to 1 × 106 cells/cm2 were prepared to further assess the effect of cell density on time-course of neuronal survival by counting the number of remaining attached neurons after 3 and 7 days in culture.
3. Neuronal responses to neurotrophic effect of NGF on neurite outgrowth and neuroprotective effect of MK-801 against glutamate-induced excitotoxity were evaluated by image analysis and MTT assay, respectively.
4. Results showed that this culture system was neuronal-enriched with a neuronal lifetime more than 35 days. Neurons survived best when seeded at a density ≥1.5 × 105 cells/cm2. Cultured neurons were capable of exhibiting sensitive responses to the effects of NGF and MK-801.
5. These findings suggest that this primary culture system provides a sensitive and powerful in vitro model for pharmacological screen and studies of neurotrophic and neuroprotective agents.