Developmental capacity of bovine oocytes matured in two kinds of follicular fluid and fertilized in vitro

This study was conducted to assess the ability of the follicular fluid derived from large and small follicles to support the in vitro oocyte maturation and subsequent fertilization and developmental capacity. Oocytes were cultured in bovine follicular fluid aspirated from small (SFF; 2–5 mm in diameter), large (LFF; 10 to 20 mm in diameter) follicles and TCM199 as a control under 5% CO2 in air. All maturation media were supplemented with 1 IU ml−1 pregnant mare serum gonadotropin. After 24 h culture, oocytes were fertilized in vitro with frozen–thawed and heparin-treated (10 μg ml−1, 15 min) bull spermatozoa and cultured in TCM199 with bovine oviductal epithelial cells (BOEC) for 7 days. Maturation of bovine oocytes cultured in LFF was inhibited and the low of male pronucleus formation was observed when compared with that of SFF (maturation rate: 69 vs. 78%; P<0.05; male pronucleus formation rate: 58 vs. 80%; P<0.05). Developmental capacity of bovine oocytes cultured in SFF was significantly (P<0.05) higher than that of LFF (15 vs. 5%), but significantly (P<0.05) lower than that of the control. There were no differences in the number of nuclei per blastocyst obtained after each treatment. These results indicate that the inhibitory action of follicular fluid on in vitro maturation, male pronucleus formation and developmental capacity of bovine oocytes is dependent on the developmental stage of the follicles from which fluid was obtained.