Culturing OVCAR-3, MeT-5A, SK-OV-3, and COV362 Cells for Proteomics

OVCAR-3 cells were maintained in RPMI-1640 (Roswell Park Memorial Institute) medium (AL162A, HiMedia) along with 10% FBS (10270, Gibco). The non-cancerous mesothelial cells, MeT-5A, were cultured in complete Medium 199 supplemented with 10% FBS, 5 μg/mL insulin, 0.5 μg/mL hydrocortisone, 2.6 ng/mL sodium selenite, 27 pg/mL β-estradiol, 10 μg/mL transferrin, 10 ng/mL hEGF, and 20 mM HEPES buffer. All the cells were maintained in a 5% ${\text{CO}}_2$ , ${37}^{\circ }$ C temperature humidified incubator. For the proteomics experiments, both 2D and 3D cultures were cultivated in defined serum-free medium in order to minimize contamination in spectrometric analysis from serum proteins. Oseltamivir phosphate (SML1606, Sigma-Aldrich) was the drug used in our study. The assay medium being defined serum-free (widely used across experimental in vitro systems⁷ (link)^,50 (link)) oseltamivir is unlikely to react with the medium. SK-OV-3 and COV362 cells were cultured in McCoy’s 5a medium (AL057A, HiMedia) with 10% Fetal Bovine Serum and DMEM (Dulbecco’s Modified Eagle Medium) medium (AL007A, HieMdia) along with 2 mm L-Glutamine, 10% FBS respectively.