Development of a Human Stomach Explant Organ Culture System to Study the Pathogenesis of Helicobacter pylori

Abstract

These studies were undertaken to define conditions under which Helicobacter (formerly Campylobacter) pylori and viable human gastric mucosa could coexist in tissue culture with the ultimate goal of developing an in vitro experimental model which could be used to study interactions between H. pylori and gastric epithelium. Antral gastric biopsies obtained at upper endoscopy were placed in culture in either CMRL-1066 or keratinocyte growth media and incubated at 37 °C in either an oxygen-enriched environment (45% O₂, 50% N₂, 5% CO₂) or a standard oxygen environment (95% air, 5% CO₂). Without selective antibiotics to suppress growth of non-H-pylori organisms, H. pylori could not be isolated from most initially positive tissue even after only 2 h in tissue culture; however, when selective antibiotics were utilized in the tissue culture media, H. pylori was isolated from 9 of 14 initially positive cases after 24–72 h in tissue culture. There was little difference in the morphology of either surface or glandular epithelium in H-pylori-negative explants between time zero and 48-hour cultures. However, H-pylori -positive explants after 48 h in tissue culture showed a significant increase in injury to both surface and glandular epithelium when compared to time zero specimens. These data demonstrate that viable H. pylori and human gastric epithelium can be maintained in explant organ culture and suggest that this gastric mucosal explant culture system may be useful in studying the significance of H-pylori infection of human gastric epithelia.