Culturing and Maintaining Diverse Cell Lines

A549 (CCL-185), MCF-7 (HTB-22), 4T1 (CRL-2539), HeLa (CCL-2), and MRC-5 (CCL-171) cells were purchased from American Type Culture Collection (ATCC). A2780 and A2780cisR cells (originally purchased from ATCC, HTB-174) were provided by W. H. Ang (Department of Chemistry, National University of Singapore). A549cisR cells were cultured as previously reported (16 , 17 (link), 58 (link)). Human breast carcinoma MCF-7 cells, human cervix carcinoma HeLa cells, human lung carcinoma A549 cells, and cisplatin-resistant A549cisR cells were cultured in Dulbecco’s modified Eagle’s medium (DMEM) containing 10% FBS and penicillin/streptomycin (100 IU ml⁻¹). Human ovarian carcinoma A2780 and A2780cisR cells were cultured in RPMI 1640 containing 10% FBS, 2 mM l-glutamine, and penicillin/streptomycin (100 IU ml⁻¹). Human lung fibroblast MRC-5 cells were cultured in MEM containing 10% FBS, 1% non-essential amino acids (NEAA), 1% l-glutamine, 1% NaPyr, and penicillin/streptomycin (100 IU ml⁻¹). In every two passages of A2780cisR and A549cisR cells, 5 μM cisplatin was added to maintain the platinum resistance. All the cells were cultured in humidified incubators at 37°C with 5% CO₂. Cell counting was performed with a hemocytometer.