The purpose of this study was to measure the culturable airborne fungi (CAF) concentrations in the underground subway stations of Seoul, Korea at two time points. This study measured the CAF concentrations in enclosed environments at 16 underground stations of the Seoul Metro in 2006 and 2013 and investigated the effects of various environmental factors, including the presence of platform screen doors, temperature, relative humidity, and number of passengers. CAF concentrations at the stations in 2006 were significantly higher than that at the same stations in 2013 (p < 0.001). Furthermore, there was a significant correlation between CAF concentration and relative humidity (r = 0.311, p < 0.05). Geotrichum and Penicillium were the predominant genera. The CAF concentrations in stations with an operating supply air were significantly higher than that in stations with no supply air (p < 0.001). Therefore, it is recommended that special attention be given to stations with clean supplied air to improve the indoor air quality of these subway stations.