A serum- and antioxidant-free primary culture model of mouse cortical neurons for pharmacological screen and studies of neurotrophic and neuroprotective agents.
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 x 10(3) to 1 x 10(6) 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 > or =1.5 x 10(5) 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.
D000818:Animals; D002452:Cell Count; D018929:Cell Culture Techniques / Q000379:methods*; D002454:Cell Differentiation / Q000502:physiology; D002470:Cell Survival / Q000187:drug effects / Q000502:physiology; D002478:Cells, Cultured / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D002540:Cerebral Cortex / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D016291:Dizocilpine Maleate / Q000494:pharmacology; D004353:Drug Evaluation, Preclinical / Q000379:methods*; D018691:Excitatory Amino Acid Antagonists / Q000494:pharmacology; D005260:Female; D005333:Fetus; D007150:Immunohistochemistry; D051379:Mice; D008954:Models, Biological; D020932:Nerve Growth Factor / Q000494:pharmacology; D009414:Nerve Growth Factors / Q000494:pharmacology*; D016501:Neurites / Q000187:drug effects / Q000648:ultrastructure; D009474:Neurons / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D018696:Neuroprotective Agents / Q000494:pharmacology*; D011247:Pregnancy
D018691:Excitatory Amino Acid Antagonists; D009414:Nerve Growth Factors; D018696:Neuroprotective Agents; D016291:Dizocilpine Maleate; D020932:Nerve Growth Factor