Guinea Pig Gastric Epithelial Cell Isolation

Primary guinea pig gastric epithelial cells were obtained as previously described [54 (link)]. Briefly, the stomach was isolated from the guinea pig, homogenized and treated with 0.25% trypsin (BIOMED-LUBLIN, Lublin, Poland) for 15 min at room temperature. The cell suspension at a density of 2 × 10⁶ cells/mL was seeded onto a 6-well plate (Becton Dickinson, Franklin Lakes, NY, USA) and cultured for 24 h (5% CO₂, 37 °C) to allow the cells to adhere. The cells were cultured in a mixture of DMEM and Ham’s F-12 media (ratio 1:1; Sigma-Aldrich, Saint Louis, Mi, USA), supplemented with 10% fetal calf serum (FCS), 1% (N-2-hydroxyethylpiperazine-N-2-ethane sulfonicacid) (HEPES), penicillin (100 U/mL), streptomycin (100 µg/mL), amphotericin B (0.025 µg/mL), l-glutamine (2 mM/mL)epidermal growth factor (Sigma-Aldrich, Saint Louis, MI, USA) 0.01 µg/mL and 0.005% dexamethasone. The guinea pig fibroblasts (CRL-1405) were obtained from the American Type Culture Collection (ATCC, Rockville, Manassas, VA, USA) The cells were routinely grown as a monolayer in complete RPMI-1640 medium cRPMI(Sigma St. Louis, MI, United States) at 37 °C in a humidified atmosphere containing 5% CO₂.Every 2–3 days, the medium was changed and the cells were passaged at 80–90% confluence.