Dulbecco modified eagle medium (dmem)

Two breast cancer cell lines (MCF-7 and MDA-MB-231) and fibroblasts as normal cell lines, which were purchased from American Type Culture Collection (ATCC, Manassas, VA, USA), were used to investigate the effects of the novel synthesized compounds. The cells were cultured in DMEM (Corning, Kennebunk, ME, USA), supplemented with FetalBovine Serum (Eurx, Gdansk, Poland) at concentration 10% and penicillin/streptomycin at 1%. Cells were cultured on 100 mm plates (Sarstedt, Newton, NC, USA) and then kept in an incubator that provided optimal growth conditions (5% CO₂, temperature: 37 °C, humidity at 90–95% level). After reaching 80% confluence, cells were detached from the bottom of the plate using 0.05% trypsin supplemented with 0.2% EDTA (Gibco, San Diego, CA, USA). Then, using a hemocytometer, the number of cells was quantified and seeded at density 5 × 105 cells per well in six-well plates (Sarstedt, Newton, NC, USA) in 2 mL of DMEM. In this research, cells that obtained 80% of confluency were used.

All cells were maintained in 10 cm dishes (Sarstedt) at 37°C and 5% CO₂. HEK293A, HEK293T-GFP and 3T3 cells were grown in DMEM (Sarstedt) supplemented with 10% FBS (Sarstedt), 1% penicillin/streptomycin + l-glutamine (Gibco), 1% sodium pyruvate (Gibco) and 50 μM β-mercaptoethanol (Gibco). CH12-F3 mouse erythroleukemia B cells were grown in RPMI 1640 (Sarstedt) supplemented with 10% FBS (Sarstedt), 1% penicillin/streptomycin + l-glutamine (Gibco), 1% sodium pyruvate (Gibco), 5% NCTC-109 (Gibco) and 50 μM β-mercaptoethanol (Gibco).