Tri gas incubator

The standard HP strains ATCC 43504 and ATCC 700392, as well as GES-1 cells, were purchased from the American Type Culture Collection (ATCC, Manassas, VA, USA). CS01 was generously granted by Professor Jing Liu (University of Shanghai for Science and Technology, Shanghai, China), and QYZ-003 and QYZ-004 were obtained from the Qingyuan Hospital of Traditional Chinese Medicine (Qingyuan, China). All HP strains were authenticated, cultured, and stored at the School of Pharmaceutical Sciences (Shenzhen, China), Sun Yat-sen University. The strains were cultured on a Columbia agar base supplemented with 5% sterile defibrinated sheep blood or in BHI broth containing 10% FBS, shaken at 150 rpm, and subsequently incubated at 37 °C in a tri-gas incubator (ESCO, Singapore) with a gas composition of 10% CO₂, 5% O₂, and 85% N₂ for a period of two to three days, as previously described [29 (link)]. GES-1 cells were cultured with RPMI 1640 medium containing 10% FBS. For co-culture, following overnight incubation of the GES1 cells on 24-well plates, HP was added to the cells at an MOI ratio of 100:1, along with varying concentrations of drugs, for a co-incubation period of 6 h before being used for subsequent experiments.

Helicobacter pylori strain ATCC 43,504 was purchased from American Type Culture Collection (ATCC, Manassas, VA, USA), ATCC 700,392 was gifted by professor Hongkai Bi (Nanjing Medical University, China), ICDC11101 was granted by professor Ping Huang (Guangzhou University of Chinese Medicine, China), LQ2# was granted by Bolaote Biotechnology (Shenzhen, China), SS1 and CS01 were granted by professor Jing Liu (University of Shanghai for Science and Technology, China). QYZ-001, QYZ-002 and QYZ-003 were obtained from Qingyuan Hospital of Traditional Chinese Medicine (Guangzhou, China). All strains used in this study were authenticated by the providers and the purity of the strain was ensured by regular morphological observation, gram staining, and biochemical reaction, and were stored at − 80 °C in 65 % BHI/ 25 % glycerol/ 10 % FBS (v/v/v). In this research, H. pylori strains were cultured in Columbia agar base supplemented with 5 % sheep blood at 37 °C in a tri-gas incubator (ESCO, Singapore) containing 10 % CO₂, 5 % O₂, and 85 % N₂ for 48 to 72 h. For liquid proliferation, strains were inoculated in BHI supplemented with 10 % FBS, vibrated at 150 rpm, under the same air environment as described above.