... After repeated washing and centrifugation to remove enzymes, the gastric glandular cells were suspended and seeded into Matrigel (R&D system, 3533-005-02, USA) and maintained in medium containing: 100 ng/mL of Wnt3A (Peprotech, 315 − 20, USA), 100 ng/mL of Noggin (R&D system, 1967-NG, USA), 500 ng/mL of R-Spondin-1 (Peprotech, 315 − 32, USA), Glutamax-I (1×) (Gibco, 35050061, USA), 1% of Penicillin/streptomycin (Gibco, 15070063, USA),100 ng/mL of FGF-10 (R&D system, 6224-FG, USA), 50 ng/mL of EGF (R&D system, 2028-EG, USA), 1×B27 (Gibco, 17504044, USA), 10 nM of Gastrin-1 (Tocris Bioscience, 3006, UK), 1 mM of N-Acetylcysteine (Tocris Bioscience, 5619, UK) during the whole cultivation and 10 µM of Y-27,632 (Tocris Bioscience, 1254, UK) for only the first 2 days. The medium was replaced three times a week [40,41]. ...