Cytotoxicity of mycotoxins evaluated by the MTT-cell culture assay.
The application of a modified colorimetric bioassay for the evaluation of the biological effects of mycotoxins is reported. Using three different monolayer cell lines (swine kidney, Madin Darby canine kidney, HeLa) the influence of nine different mycotoxins on the cellular methylthiazoltetrazolium (MTT)-cleavage activity was evaluated. The yellow tetrazolium salt MTT is converted by mitochondrial dehydrogenases of metabolically active cells to an insoluble purple formazan product, which was then solubilized with dimethylsulfoxide. The optical density of this homogeneous solution was suitable for a precise spectrophotometric measurement by a plate reader at a wavelength of 510 nm. Nine mycotoxins were simultaneously tested in all three cell lines, from which the swine kidney cell line proved to be the most sensitive. The effects of additional 35 mycotoxins were therefore tested using swine kidney monolayers as target cells. A total of 28 toxins of the 44 mycotoxins tested proved to be cytotoxic in the MTT-bioassay. Most of them belong to the group of trichothecene mycotoxins. Concentrations ranged between 0.01 micrograms and 100 micrograms/ml of cell culture medium. The MTT cleavage assay was found to be a quick (24 hours) and easy to perform system for the evaluation of the biological activity of many different mycotoxins and may also provide a useful tool for the testing of a large variety of sample materials.
Hanelt
M
M
Institute of Medical Microbiology, Infectious and Epidemic Diseases, Veterinary Faculty, Ludwig-Maximilian-University Munich, Germany.
Gareis
M
M
Kollarczik
B
B
D000818:Animals; D000935:Antifungal Agents / Q000633:toxicity; D016923:Cell Death / Q000187:drug effects; D002478:Cells, Cultured / Q000187:drug effects; D004285:Dogs; D006367:HeLa Cells; D006801:Humans; D007668:Kidney / Q000166:cytology / Q000187:drug effects; D009183:Mycotoxins / Q000633:toxicity*; D013552:Swine; D013778:Tetrazolium Salts; D013844:Thiazoles
D000935:Antifungal Agents; D009183:Mycotoxins; D013778:Tetrazolium Salts; D013844:Thiazoles; C022616:thiazolyl blue