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Aim: The aim was to assess the effects of fetal bovine serum (FBS) and estrus buffalo serum (EBS) on in vitro maturation rate of oocytes in buffalo. Materials and Methods: Maturation rate of oocytes was assessed in two maturation media supplemented with 20% FBS and EBS. Oocytes maturation rate was evaluated on the basis of cumulus cell expansion and extrusion of polar body after 24 h of in vitro culture in CO2 incubator. Results: The average percentage of in vitro matured oocytes in FBS was 83.80%, and EBS was 77.45%, respectively. The results revealed a significant (p<0.05) increase in maturation rate of oocytes in FBS than EBS. Conclusion: Buffalo oocytes were better in vitro matured in FBS than EBS.