To simplify the procedure for superovulation in the rhesus monkey, this study was designed using polyvinylpyrrolidone (PVP) solution as a solvent for gonadotropins. Thirty-five cycling females (aged 5-8 years old) were divided into six groups during the breeding season (November- February). The groups were as follows: Group I, animals received twice-daily 35 IU recombinant human follicle-stimulating hormone (rhFSH) dissolved in 0.5 ml saline for 8 days as the control; Groups II and III, animals received single-daily 35 IU and 17 IU rhFSH in 0.5 saline, respectively, for 9 days; Groups IV, V and VI, received single-daily injection of 35 IU rhFSH, 17 IU rhFSH and 8.5 IU rhFSH dissolved in 0.5 ml 30% PVP (w/v) solution, respectively, for 9 days. After human chorionic gonadotropin was administered to induce the nuclear maturation of oocytes, oocytes were retrieved and the development competence of recovered oocytes treated with in vitro fertilization were observed. The plasma concentrations of follicle-stimulating hormone and ovarian responses were monitored during the treatment. The results showed that the number of recovered oocytes and the in vitro developmental competence of mature oocytes was equivalent among monkeys when treated with a single-daily treatment of 17 and 35 IU rhFSH with PVP preparation in Groups IV and V compared with the twice-daily 35 IU rhFSH treatments received by Group I. However, almost all animals in Groups II, III and VI responded poorly to corresponding stimulations. These findings indicate that a single-daily low dose of rhFSH dissolved in PVP solution can induce the satisfactory ovarian responses in rhesus monkeys. This has the potential to reduce treatment distress, stress to the animals, the labor of the operator as well as the amount of rhFSH used in ovarian stimulation, compared with traditional superovulation methods.