Retinoic acid (RA), a vitamin A metabolite, regulates cellular growth, embryonic morphogenesis and differentiation. In the present study, the effect of various doses of RA (0, 1, 3 and 5 μM) on in vitro maturation (IVM) of water buffalo oocyte was determined. Nuclear maturation and cumulus expansion of the oocytes were enhanced by RA. Cumulus expansion rates in the treatment groups at 1 (80.67%), 3 (80%) and 5 (86%) μM were significantly different compared with the control (72.02%) and the vehicle (67.50%). The percentage of oocytes reaching the metaphase II stage was significantly increased (P<0.05) in the RA-treated group compared with the control (67.97%) and vehicle (66.46%), with 5 μM RA dose (84.47%) exhibiting the highest rate. RA supplementation improved cleavage rate. Treatment with 5 μM RA significantly improved blastocyst formation rate (38.88%) compared with the control and the vehicle. However, there was no significant difference between the control and RA-treated groups in the number of cells of the resultant blastocysts. The results demonstrated that RA enhanced IVM of oocytes to positively influence the development of water buffalo embryos after in vitro fertilization.