title: Hydrogen peroxide-induced neurotoxicity in cultured cortical cells grown in serum-free and serum-containing media.
abstract: To compare different culture conditions for neuroprotection assays in cultured cortic neurons, we evaluated cell viability after H2O2 exposure in cells cultured with standard N2 and with the enriched B-27 developed by GIBCO, both serum-free supplements. The following additives/associations were compared: N2 (+N2), B-27 (+B-27), 10% FBS (+FBS), 1% FBS in combination with N2 (FBS/N2) or N2 supplement preceded by an 1 hour precoating with 10% FBS (N2 + precoated). Our data demonstrated that B-27 is as efficient as 10% FBS to support neuronal growth for more than a week. As shown by phase-contrast optics cells grown in N2 started degenerating within 24-48 hours although measurable absorbance was seen with MTT. The pre-coating procedure failed to modify substantially cell viability as compared with N2 alone. Dose-response curves for H2O2 to induce neuronal apoptosis were almost identical for B-27 and serum supplemented samples. Catalase (100 U/ml) or vitamin E (200 microM) prevented cell death in both culture conditions. Our results indicate that DMEM/B-27 provides a serum-free cell culture environment that allows neurons to grow with optimal cell viability, comparable to that obtained with serum. We conclude that this culture condition reveals as a useful tool to test the efficacy of neuroprotectants when a serum free medium is required.
authors.affiliation: Laboratorio de Neurociencias, Centro de Investigaciones Médicas Albert Einstein-Fundación CIMAE, Buenos Aires, Argentina.
authors.forename: K C
authors.initials: KC
authors.lastname: Ricart
authors.forename: M L
authors.initials: ML
authors.lastname: Fiszman
mesh_terms: D000818:Animals; D001769:Blood; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D002540:Cerebral Cortex / Q000166:cytology / Q000187:drug effects* / Q000502:physiology; D003470:Culture Media / Q000494:pharmacology*; D016895:Culture Media, Serum-Free / Q000494:pharmacology; D006861:Hydrogen Peroxide / Q000494:pharmacology*; D051379:Mice; D009474:Neurons / Q000187:drug effects / Q000502:physiology; D009498:Neurotoxins / Q000494:pharmacology*; D016877:Oxidants / Q000494:pharmacology*; D018384:Oxidative Stress
chemical_list: D003470:Culture Media; D016895:Culture Media, Serum-Free; D009498:Neurotoxins; D016877:Oxidants; D006861:Hydrogen Peroxide