title: A serum- and antioxidant-free primary culture model of mouse cortical neurons for pharmacological screen and studies of neurotrophic and neuroprotective agents.
abstract: 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.
mesh_terms: 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
chemical_list: D018691:Excitatory Amino Acid Antagonists; D009414:Nerve Growth Factors; D018696:Neuroprotective Agents; D016291:Dizocilpine Maleate; D020932:Nerve Growth Factor