The survival of six clinical isolates of Helicobacter pylori at room temperature was investigated after suspension in five different media: brucella broth with 5% lysed horse blood (BBLH), phosphate-buffered saline (PBS), 20% glucose, Stuart medium, and PBS with 10% Fildes enrichment (PBS-F). Only in BBLH and PBS-F no decrease in mean bacterial numbers was observed during the 24-h study period. No H. pylori isolates could be cultured from Stuart medium after 7 h of incubation. In contrast, the recovery rates in PBS-F or Stuart medium of H. pylori isolates from gastric tissue specimens collected from 19 H. pylori-positive patients were not significantly different even after a delay of culture of up to 24 h. Our data show that the medium composition is not critical for the survival of H. pylori within gastric tissue specimens.