Development of a fish in vitro cell culture model to investigate oxidative stress and its modulation by dietary vitamin E

When cultured in Dulbeccoʹs minimal essential medium the established epithelioma papulosum cyprini cell line from carp was found to be vitamin E-deficient due to the very low level of vitamin E in the medium and the foetal calf serum used as supplement. The toxicity of oxidative stressors to this cell line was evaluated by means of the neutral red cytotoxicity assay and it was found that an organic hydroperoxide, t-butylhydroperoxide was extremely cytotoxic and that the redox-cycling agents diquat and menadione were less toxic. When grown under vitamin E supplementation (25 μM), the toxicity of these chemicals was reduced by at least an order of magnitude in concentration demonstrating the protective effect of vitamin E. These data show the importance of vitamin E status for interpretation of in vitro and in vivo data and that this in vitro system is useful for mechanistic studies.