Model cb160

We extracted H. pylori strains containing the preservation solution at −80°C. Standard strains 26,695, NSH57, MSD132, and G27 were donated by Bi Hongkai from the Nanjing Medical University, and clinical strains HPBS001–HPBS016 were isolated at our laboratory. H. pylori strains were cultured on the Columbia blood agar plate (OXOID, UK) or in the brain-heart infusion (BHI, OXOID, UK) broth medium containing 10% serum (Pingrui, China) and placed under microaerophilic (85% nitrogen, 5% oxygen, 10% carbon dioxide; model CB160; Binder, Germany) conditions at 37°C for 3 days. Bacterial species other than H. pylori were cultured on nutrient agar or Luria–Bertani plates at 37°C for 1–2 days. Supplementary Table S1 shows the information of Staphylococcus aureus and other information.

BMCs were collected from femurs and tibias of adult rats, flushed with sterile phosphate-buffered saline (PBS; Carl Roth GmbH, Karlsruhe, Germany), incubated in red blood cell lysis buffer (Sigma, Welwyn Garden City, UK) and cultured in Iscove’s Modified Dulbecco’s Medium (IMDM, Pan Biotech, Aidenbach, Germany) supplemented with 20% Fetal Bovine Serum (FBS, Biochrom AG, Berlin, Germany), 100 IU mL⁻¹ penicillin and 100 mg mL⁻¹ streptomycin (Corning, Corning, NY, USA). The initial culture medium was changed at passage 1 to Dulbecco’s Modified Eagle’s Medium–high glucose (DMEM, Sigma–Aldrich, Schaffhausen, Switzerland), with 10% FBS, 100 IU mL⁻¹ penicillin and 100 mg mL⁻¹ streptomycin. Cells were cultured under aseptic conditions using sterile and RNAse/DNAse free tissue culture-treated plastic ware (Corning, NY, USA) at 37 °C and 5% CO₂ in a humidified incubator (Model CB160, Binder, Tuttlingen, Germany). The medium was replaced every two days, and upon 80% confluence, cells were trypsinized, pooled, and subcultured until passage 2. In order to preserve the different populations of the BMCs, cells were collected at passage 2, and no other selection was performed.
For in vivo study, each pool was obtained from 10 male Lewis rats (mean weight of 150 g). For the in vitro study, each pool of BMCs was prepared from 3 male Lewis rats (mean weight of 150 g).