PROBLEM: The aim of this study was to determine the presence and concentration of interleukin IL‐6, IL‐8, and granulocyte‐macrophage‐colony‐stimulating factor (GM‐CSF) in pre‐ovulatory ovarian follicular fluid (FF) of patients undergoing controlled ovarian hyperstimulation for intracytoplasmic sperm injection (ICSI) therapy on one hand, and to find out whether these cytotokine concentrations could be used as a predictive parameter for ICSI outcome. DESIGN: The levels of IL‐6, IL‐8, and GM‐CSF were measured from women that underwent ICSI therapy and the results were compared between the patients who became pregnant after ICSI and those who did not. MATERIAL AND METHODS: FF was obtained from 84 women participating in our ICSI program. The patients underwent ovarian hyperstimulation with human menopausal gonadotropin after they had been treated with gonadotropin‐releasing hormone agonist (GnRH‐a) for pituitary down‐regulation. The concentration of IL‐6, IL‐8, and GM‐CSF was measured by the highly sensitive enzyme‐linked immunosorbent assay method. RESULTS: Twenty‐eight women became pregnant (group I) and the other 56 (group II) did not. The main concentrations of IL‐6, IL‐8, and GM‐CSF in FF of patients who became pregnant were 5.4±2.9, 207.9±118.3, and 1.01±0.8 pg/mL, respectively, and the corresponding values in patients who did not become pregnant were 7.0±5.1, 230.8±183, and 1.34±0.09 pg/mL, respectively. No statistical difference was found between the two groups with regard to the age of patients, IL concentration in FF, mean number of retrieved oocytes, mean number of mature oocytes (metaphase II), and the fertilization rate. CONCLUSION: IL‐6, IL‐8, and GM‐CSF were found in most FF investigated either from patients who became pregnant after ICSI treatment or from those who did not. Furthermore, no significant differences were found between the IL‐6, IL‐8, and GM‐CSF concentrations in both groups. The mean oocyte retrieval, fertilization, and cleavage rates were not significant between the patients who became pregnant and those who did not. Therefore, the IL‐6, IL‐8, and GM‐CSF concentrations in pre‐ovulatory FF cannot be used as a reliable prognostic index for fertilization ability, developing potential, and implantation probability.