Five grains of iridin, mixed with a little bile and water, and placed in the duodenum, very powerfully stimulate the liver

MCF-7 and T47D human breast cancer cells were used for activity evaluation. Cells were seeded into 96-well tissue culture plates in 5% DCC-treated, FBS-supplemented RPMI phenol red-free medium at a density of 1 x 10^4 cells/well. Test compounds were added in DMSO solution (control contained 1% DMSO) and incubated at 37 °C with 5% CO2 for 96 h. In all experiments, serial dilutions of estradiol were added as a positive control. To evaluate relative cell concentrations, Alamar Blue reagent was used. After 3 h, fluorescence was measured at 590 nm with excitation at 530 nm using a FL500 spectrophotometer