Cytotoxicity Assessment of Cancer Cell Lines

MDA-MB-231 (breast cancer), A549 (lung cancer), and normal human fibroblasts cell lines were cultured in full medium containing DMEM supplemented with 10% FBS, 1% antimycotic at 37 °C, and 5% CO₂ in 75 cm² tissue culture flasks. For the cytotoxicity assessment, cells were trypsinized (Trypsin 0.05%/0.53 mM EDTA) and reaction was stopped by adding full medium to collect all cells. Then, cells were seeded (2 × 10³ cell/well in 200 µL of medium) into 96-well plates (Gibco, NY, USA) for 24 h before being treated with different concentrations of pure oils, drugs, and/or their different formulations for another 72 h [28 (link)].