title: Lasting effect of preceding culture conditions on the susceptibility of C6 cells to peroxide-induced oxidative stress.
abstract: The aim of the present study was to investigate the influence of the maintenance culture conditions on the competence of C6 rat glioma cells to cope with peroxide-induced oxidative stress. C6 cells were maintained either in Ham's nutrient mixture F-10 supplemented with 15% horse serum and 2.5% foetal bovine serum (FBS) or in Dulbecco's Modified Eagle's Medium (DMEM) supplemented with 5% FBS. The differently cultured cells were exposed under identical conditions to hydrogen peroxide (H₂O₂) and cumene hydroperoxide (CHP) in serum-free DMEM. The cells maintained in high serum Ham's F-10 medium (1) were less sensitive towards the cytotoxic action of both peroxides (EC₅₀-values: H₂O₂: 193 ± 23 μM; CHP: 94 ± 16 μM) than the cells maintained in low serum DMEM (EC₅₀-values: H₂O₂: 51 ± 10 μM; CHP: 27 ± 11 μM), (2) eliminated the peroxides (initial concentration: 100 μM) with higher rates (H₂O₂: 56 ± 5.5 vs. 32 ± 2.7, CHP: 32 ± 6 vs. 3.4 ± 0.6 nmol/min mg protein), (3) contained more glutathione (30 ± 2.5 vs. 14 ± 1.1 nmol/mg protein) and (4) owned a higher glutathione peroxidase activity (28 ± 3.4 vs. 9.5 ± 0.8 mU/mg protein). Glutathione reductase and catalase activities were not affected. These results demonstrate that the preceding culture conditions have a lasting effect on the susceptibility of cultured cells to oxidative stressors like peroxides. As cause for these differences a dissimilar supply of the cells with serum born antioxidants like selenium and α-tocopherol is discussed.
authors.affiliation: Institute for Toxicology and Pharmacology for Natural Scientists, University Hospital Schleswig-Holstein, Campus Kiel, Brunswiker Str. 10, 24105 Kiel, Germany.
authors.forename: Sibylle
authors.initials: S
authors.lastname: Brenner
authors.forename: Michael
authors.initials: M
authors.lastname: Gülden
authors.forename: Edmund
authors.initials: E
authors.lastname: Maser
authors.forename: Hasso
authors.initials: H
authors.lastname: Seibert
mesh_terms: D000818:Animals; D001555:Benzene Derivatives / Q000633:toxicity; D002374:Catalase / Q000378:metabolism; D018929:Cell Culture Techniques / Q000379:methods*; D045744:Cell Line, Tumor; D005978:Glutathione / Q000378:metabolism; D005979:Glutathione Peroxidase / Q000378:metabolism; D006861:Hydrogen Peroxide / Q000633:toxicity; D016877:Oxidants / Q000633:toxicity*; D018384:Oxidative Stress; D010545:Peroxides / Q000633:toxicity*; D051381:Rats
chemical_list: D001555:Benzene Derivatives; D016877:Oxidants; D010545:Peroxides; D006861:Hydrogen Peroxide; D002374:Catalase; D005979:Glutathione Peroxidase; D005978:Glutathione; C007164:cumene hydroperoxide