The reproductive biology of the domestic dog is unique among mammalian species; because of this, in vitro maturation (IVM) rate is still very low compared with other domestic animals in spite of attempts at improvement. The aim of this study was to consider the use of recombinant human gonadotropins as a replacement for pituitary gonadotropins for IVM of dog oocytes. A total of 845 cumulus-oocyte complexes were used in this study. To determine the effects of human recombinant gonadotropins, maturation medium was supplemented with two different concentrations (0.05 or 0.1 IU/mL) of pituitary (pFSH, pLH) and human recombinant (rhFSH and rhLH) gonadotropins. After the IVM period, the maturation rate of the oocytes was investigated under an epifluorescence microscope. Our findings showed no significant difference in maturation rate using either pituitary or human recombinant gonadotropin groups (P > 0.05). Applying 1.0 IU human recombinant gonadotropin caused the lowest maturation rate (34.57%; P < 0.05). In conclusion, recombinant human gonadotropins could be applied for IVM of dog oocytes. Moreover, 0.05 IU/mL rhFSH and rhLH can be successfully used in place of biologically derived hormones.