Soy isoflavones are phytoestrogens which have been associated with several health benefits. In the present study, we report the production of isoflavones in a collection of 40 strains of soya cell cultures established in 1975. A large variability in the isoflavone composition was observed and high-producing strains, with an isoflavone content of up to 46.3 mg g(-1) dry wt., were found. In comparison with soybeans, many callus strains had a higher isoflavone concentration (10-40 times) and a different ratio of genistin to daidzin forms. The highest producing strain was transferred to liquid medium in an Erlenmeyer flask and in a 10 l stirred-tank bioreactor where high isoflavone content (7% dry wt.), concentration (880 mg l(-1)) and a maximum productivity estimated to 60 mg l(-1) d(-1) were obtained. We further studied the estrogenic activity of pure compounds compared to plant cell culture extracts in the estrogen-responsive human endometrial Ishikawa cell line. Estrogen was confirmed to be 1000-10,000 times more active than isoflavones. The estrogenic activity of the extracts correlated to their isoflavone content. The activity of the malonyl isoflavones, assessed here for the first time, was lower than the aglycones. Taken together, these results suggest that soya cell cultures can be used as an alternative source to soybeans to provide high concentrations of bioactive isoflavones.