Comparison of the cytotoxicity of the MEIC reference chemicals measured in protein free and in complete culture medium

In order to investigate if a protein free cytotoxicity assay could improve the prediction of human acute toxicity, the cytotoxicity of 40 MEIC reference chemicals was measured by the neutral red uptake inhibition after 24 h in protein free culture medium on rat hepatoma-derived Fa32 cells. The results were compared with the corresponding values obtained in complete culture medium, including 10% fetal calf serum. Potassium cyanide, arsenic trioxide, mercuric chloride, hexachlorophene and pentachlorophenol were much more cytotoxic in PF medium, as was the case to a lower extent for 16 other chemicals. The cytotoxicity of 8 chemicals was only changed to a limited extent when tested in PF medium, suggesting that serum proteins do not strongly interact with their cytotoxicity. Eleven other chemicals were less cytotoxic in PF medium, maybe because of too poor physiological conditions. Although a large number of differences in cytotoxicity were observed in function of the medium used for the assay, a good correlation was observed between both series of data (r2 = 0.946). The correlation between the cytotoxicity in PF medium and the human acute toxicity is lower (r2 = 0.647) than that in complete medium (r2 = 0.746). The results show that further research is necessary in order to improve the in vitro/in vivo correlations by introducing protein-dependent considerations.