Mcf 7

The human breast adenocarcinoma cell line (MCF7) was obtained from National Centre for Cell Science (NCCS), Pune and grown in Eagles Minimum Essential Medium containing 10% fetal bovine serum (FBS). The cells were maintained at 370֩ C, 5% CO₂, 95% air and 100% relative humidity. Maintenance cultures were passaged weekly, and the culture medium was changed twice a week. The human embryonic kidney cell line (HEK 293) was obtained from National Centre for Cell Science (NCCS), Pune and grown in Eagles Minimum Essential Medium containing 10% fetal bovine serum (FBS). The cells were maintained at 370֩ C 5% CO2, 95% air and 100% relative humidity. Maintenance cultures were passaged weekly, and the culture medium was changed twice a week.

Human breast cancer cell lines (MCF-7) were purchased from the National Centre for Cell Sciences (Pune, India). These cells are regularly cultured according to the procedure described by Shagufta et al. and Gupta et al.[20 (link)21 (link)] For culturing the MCF-7 cells, DMEM (pH 7.4) was used which contains streptomycin (100 U/mL), penicillin (200 mg/mL), and gentamicin (50 mg/mL) supplemented with 10 mM HEPES and 10% FBS at 37°C at a humidified atmosphere of 5% CO₂ incubator in T-25 tissue culture flask (TPP, Switzerland). Before performing the experiments, the cells were trypsinized which were obtained from a confluent flask and further cultured for 4 days. For the initial 2 days, DMEM phenol red free was used which contains 10% charcoal-stripped FBS to preculture the cells.[22 (link)] Subsequently, the cells were exposed to ligand (Steviol) for 48 h. The number of cells required for analyzing cytotoxicity and the exposure of cells with different concentrations of ligands has been described individually below.