Sensitivity of cultured human and mouse fibroblasts to trichothecenes

Abstract

Cultured human diploid fibroblasts and a cloned permanent line of Swiss mouse fibroblasts (3T3) have been adapted to a biological assay of trichothecenes and other toxic fungal metabolites both in purified form and as crude extracts. The assays are carried out in multi-well tissue culture trays with serial dilutions of toxins in 200 microL volumes of medium. Cells are cultured with the toxins for 3-4 days, fixed, and stained, providing a permanent record of results. The sensitivity of the assay ranges from 0.1 ng/mL (20 pg/assay) for T-2 toxin on mouse fibroblasts to no detectable toxicity at 5 micrograms/mL for cis-zearalenone and verrucarol. Human skin fibroblasts exhibit patterns of relative sensitivities similar to that observed with mouse fibroblasts, but the absolute sensitivities are much greater with mouse fibroblasts because they can be cultured at much lower cell densities.