In vitro alternatives to whole animal testing. Comparative cytotoxicity studies of divalent metals in established cell lines derived from tropical and temperate water fish species in a neutral red assay

A simple vital dye (neutral red) uptake method for investigation of the acute toxicity of seven divalent metal cations to cell lines derived from the turbot (TF) and bluegill (BF-2) was evaluated. The method was extremely reproducible between replicate experiments on a week to week basis as well as being reproducible between different laboratories. Whilst there were some species-dependent differences in the relative toxicities of the metals there were good correlations between toxicity and the chemical softness (Σp) of the metal ions and also between in vivo and in vitro toxicities. These studies demonstrate that this simple and relatively inexpensive technique carried out in 96-well microplates may be extremely useful for evaluation of structure-toxicity relationships between related groups of compounds.