ALTEX :: Alternatives to Animal Experiments

1990, Volume 2

Tissue Culture Tests as an Alternative to the Uterotrophic Assay in the Detection of Estrogenic Active Compounds

Ulrich Mayr

Institut für Tierernaehrung der Universitaet Hohenheim, D-Stuttgart

SUMMARY

The bases for two biological systems which ascertain estrogenic activity were worked out. The primary aim was to assess animal foodstuff. The first system, the measurement of estrogen-specific proteins in MCF-7 cells (human breast cancer cells), has been used for a quantitative estimation of 17β-estradiol, the mycoestrogen zearalenone and five phytoestrogens. Estimates of the relative estrogenic potentials of these substances using the MCF-7 test yields results similar to those of the uterotrophic assay. Analyses of cereal samples artificially contaminated with zearalenone showed that MCF-7 cells are suitable for examining not only purified compounds, but also biological material. A quantitative estimation of added zearalenone was possible in each case. In principle, then, the MCF-7 system can be suitably introduced into agronomic practice, for example in the examination of animal foodstuff.

The second test system is based on genetically transformed mouse cell cultures. The synthesis of a bacterial enzyme (chloramphenicol-acetyltransferase, CAT) is used to measure hormone activity. This system was tested with zearalenone. Both test system are extremely sensitive and superior to the uterotrophic assay with mice and rats, concerning the accuracy of measurement and reproducibility. They can replace this commonly used bioassay in most cases--both in agronomic and in pharmaceutical research.

Ways of simplifying the experimental procedures are discussed.

Keywords: estrogenic activity, breast cancer cells, transgenic mouse cell culture, uterotropic assay