The further development of rainbow trout primary epithelial cell cultures as a diagnostic tool in ecotoxicology risk assessment
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The use of short-term cytotoxicity assays for the initial screening of chemicals not only aids in establishing priorities for the selection of chemicals that should be tested in vivo, but also decreases the time in which potential toxicants can be valued. Rainbow trout primary skin epithelial cell cultures are one such assay. Rainbow trout primary skin cell cultures contain two cell types, keratinocytes and goblet mucus cells. Two aquatic pollutants, copper and prochloraz were screened using this cell system. The influence of media composition on the effects of the aquatic pollutants was also studied by testing the chemicals in both serum-containing and serum-free medium and the morphological changes that occurred within the cell cultures recorded. The concentration of copper that causes a reduction of 90% in the residual of day 3 growth of the primary cell culture system was found to be approximately 10 fold more than that of prochloraz. Prochloraz was found to cause a greater reduction in growth area when added to the primary cell culture system in serum-free medium than in serum-containing medium. Copper, in contrast, was found to exert reduced toxicity when added to the test cultures in serum-free medium compared with addition in serum-containing medium. Prochloraz was found to kill the epithelial cells by a process of necrosis. Copper, was found to kill the epithelial cells by both necrosis and apoptosis in a ratio of 2:1. It was also observed that as the dose of both chemicals increased, the number of goblet cells contained in the cell cultures decreased. A PAS stain was carried out to determine if the goblet cells were exocytosing their contents onto the cell culture surface. It was found that as chemical exposure increased the number of cells expressing positivity for mucus also increased. The results of this study add further evidence to support that primary cell cultures are a very appropriate model for toxicity risk assessment.
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