The main objective of this study was to evaluate the effect of supplementing maturation media with, retinol (10 µmol/ml), ascorbic acid (100 µmol/ml) and α-tocopherol (100 µmol/ml), on cumulus cell expansion and in vitro nuclear maturation of buffalo oocytes. Oocytes were collected from ovaries taken from slaughterhouses. Only compact oocytes were used for in vitro maturation by TCM-199 medium supplemented with different vitamin levels for 22 h in CO2 incubator at 38.5 ^°C, 5% CO2 and 95% humidity.  Maturation rate was determined in terms of degree of expansion of cumulus cells and oocytes at metaphase-II (nuclear maturation). Results revealed that all supplements yielded higher percentages of oocytes showing full expansion of cumulus cells (77.2, 69.4 and 69.2% for tocopherol, ascorbic acid and retinol, respectively) as compared to control medium (60.2%), but the differences were not significant. Maturation rate in term of percentage of oocytes arrested at anaphase-I, telophase-I and metaphase-II was significantly higher for media supplemented with retinol, tocopherol and ascorbic acid, being 83.5, 82.2 and 81.94% as compared to 68.54% in control medium, respectively. Conclusively, the present results may suggest that the supplementation of maturation medium (TCM-199) with retinol, ascorbic acid and α-tocopherol as a non enzymatic antioxidants had a beneficial effect on the cumulus cell expansion and nuclear maturation of buffalo oocytes. Retinol supplementation yielded the best results.