Cultivation experiments targeting chemolithoautotrophic microorganisms were performed using subseafloor basaltic cores (the deepest sample is from 315 meters below seafloor [mbsf] and overlying sediment cores (the deepest sample is from 91.4 mbsf) from North Pond on the western flank of the Mid-Atlantic Ridge. The cores were recovered by the R/V JOIDES Resolution during Integrated Ocean Drilling Program Expedition 336. Different bacteria were grown under different media and temperature conditions. In the enrichment cultures of the basaltic cores under aerobic conditions, frequently detected bacteria at 8°C and 25°C were members of the genera Ralstonia (the class Betaproteobacteria) and Pseudomonas (Gammaproteobacteria), whereas members of the genera Paenibacillus (Bacilli) and Acidovorax (Betaproteobacteria) were conspicuous at 37°C. Bacillus spp. (Bacilli) were outstanding at 37°C under anaerobic conditions. In the enriched cultures of the sediment cores, bacterial growth was observed at 15°C but not at 37°C, and the bacteria detected at 15°C mostly belonged to gammaproteobacterial genera such as Pseudomonas , Halomonas , and Marinobacter . All of the bacteria detected in this study were enriched only, and subcultivation of the enriched cultures in the respective original media did not succeed. The presence of hydrogenotrophic methanogens was examined by a culture-dependent or a culture-independent analysis in the basalt and sediment cores but was not proven. A fungal isolate was obtained from a single basaltic core an d belonged to the genus Exophiala of the order Chaetothyriales.