Mem vit

The studies focused on elucidating human gastric cancer cells: daunorubicin—sensitive EPG85-257P and daunorubicin resistant EPG85-257 RDB, obtained as a kind gift from Prof. Herman Lage. EPG85-257 RDB cell line indicates resistance to daunorubicin and overexpression of MDR1 (known as P-gp) mechanism involvement [1 (link)]. Gastric cell lines were grown in Leibovitz L-15 medium (Sigma-Aldrich, Poznan, Poland) supplemented by 10% fetal bovine serum (FBS, Lonza, Celllab, Warsaw, Poland), 1% of antibiotics (penicillin/streptomycin, Sigma-Aldrich, Poland), 1 mM ultraglutamine (Sigma-Aldrich, Poznan, Poland), 6.25 mg/L fetuin (Sigma-Aldrich, Poland), 2.5 mg/mL transferrin (Sigma-Aldrich, Poznan, Poland), 0.5 g/L glucose, 1.1 g/L NaHCO₃, 1% minimal essential vitamins (Mem-Vit, Sigma-Aldrich, Poznan, Poland). The cells were maintained in a humidified atmosphere at 37 °C and 5% CO₂ (SteriCult, Thermo Scientific, Alab, Warsaw, Poland). For the experiments, the cells were removed by Trypsin-EDTA 0.25% solution (Sigma-Aldrich) and washed with PBS (phosphate-buffered saline) (BioShop, Lab Empire Poland, Rzeszow, Poland).

Two gastric adenocarcinoma cell lines were used: the EPG85-257 P – sensitive and EPG85-257 RDB – daunorubicin-resistant cell line. Human gastric cancer cells were obtained as a kind gift from Prof. Herman Lage. The EPG85-257 RDB cell line reveals the overexpression of MDR1 and the resistance to daunorubicin [37 (link)]. Both cell lines were cultured in Leibovitz L-15 medium (Sigma-Aldrich, Poznan, Poland) supplemented by 10% fetal bovine serum (FBS, Lonza, Poland), 1% antibiotics (penicillin/streptomycin, Sigma-Aldrich, Poznan, Poland), 1 mM ultraglutamine (Sigma-Aldrich, Poznan, Poland), 6.25 mg/L fetuin (Sigma-Aldrich, Poznan, Poland), 2.5 mg/mL transferrin (Sigma-Aldrich, Poznan, Poland), 0.5 g/L glucose, 1.1 g/L NaHCO₃, and 1% minimal essential vitamins (Mem-Vit, Sigma-Aldrich, Poznan, Poland). Cell cultures were cultured as a monolayer on a 25 and 75 cm² plastic flask (Sarstedt, Germany) maintained in a humidified atmosphere at 37 °C and 5% CO₂, and detached for the experiments by trypsinization (trypsin 0.025% and EDTA 0.02% solution, Sigma-Aldrich, Poznan, Poland). Cells were passed every 2–3 days and a day before the experiment.