Ham s f 12 medium

The human GC cell lines HGC27 and AGS were purchased from the Cell Bank of Type Culture Collection of the Chinese Academy of Sciences (Shanghai, China), and the human GC cell line MGC803 and human gastric epithelium cell line GES-1 were obtained from the Key Laboratory for Tumor Precision Medicine of Shaanxi Province (Xi’an, China). HGC27 and MGC803 cells were cultured in the RPMI 1640 medium (Sigma-Aldrich, WI, USA), AGS cells were maintained in Ham’s/F-12 medium (Procell, Wuhan, China), and GES-1 cells were grown in Eagle’s minimum essential medium (DMEM, Sigma-Aldrich, WI, USA), containing 10% fetal bovine serum (Biological Industries, Israel) and 1% penicillin/streptomycin (New Cell & Molecular Biotech, Suzhou, China). All cells were incubated in a water-saturated atmosphere of 5% CO₂ at 37 °C and were collected at the peak of the logarithmic growth phase for experiments.

This study obtained the human GC cells (AGS, HGC27, and MKN28), and normal human gastric epithelial Ges-1 cells from the Institute of Biochemistry and Cell Biology (Shanghai Institute of Biological Sciences, Chinese Academy of Sciences). AGS cells were cultured in Ham’s F12 medium (Procell Life Science & Technology Co., Ltd., Wuhan, China) with 10% fetal bovine serum (FBS; Gibco, Shanghai, China), streptomycin (50 ug/ml), and penicillin (50 U/ml) (Solarbio, Beijing, China). With 10% FBS (Gibco) and penicillin–streptomycin (Solarbio), HGC27, and MKN28 cells were cultured in RPMI-1640 supplemented (Gibco). Ges-1 cells were cultured in DMEM high glucose medium (Gibco), including 10% FBS (Gibco) and penicillin–streptomycin (Solarbio). In line with specific protocols, we maintained cells within the humid incubator under 37°C and 5% CO₂ conditions.