The present research investigated the effects of eight different culture media(numbered as A through H) on the nutritional composition, polyphenol contents and antioxidant activity(DPPH and ABTS free radical scavenging activities, and reducing power) of Pleurotus ostreatus. Results showed that Pleurotus ostreatus from various culture media presented differences in their nutritional composition, polyphenol contents and antioxidant activities. The highest contents of ash, sugar and dietary fiber were detected in Pleurotus ostreatuses cultured with medium H, and the highest contents of water, protein and fat were detected when the edible mushroom was cultured with media C, F and B, respectively. Differences in all investigated mineral elements were seen in the Pleurotus ostreatus cultured with various culture media, and some of them even showed great differences. Six fatty acids were found in different amounts among eight samples of Pleurotus ostreatus, together with differences in polyphenol contents. Polyphenols reached the highest level when medium E was used. The antioxidant activity was enhanced as the concentration of polyphenols(for both free and bound forms) increased. However, the differences in each antioxidant activity of Pleurotus ostreatus cultured with various culture media varied with the concentration of polyphenols.