Effects of follicular fluid on fertilization and embryonic development of bovine oocytes in vitro

• Published in: Theriogenology 1998-04, Vol.49 (6), p.1103-1112
• Main Authors: Choi, Y.H, Takagi, M, Kamishita, H, Wijayagunawardane, M.P.B, Acosta, T.J, Miyazawa, K, Sato, K
• Format: Article
• Language: English
• Subjects: Animals; BLASTOCYST; BODY FLUIDS; BOVIN; bovine; CATTLE; Cattle - embryology; Cattle - physiology; CELL DIVISION; CLEAVAGE; Coculture Techniques - veterinary; CULTIVO DE EMBRIONES; CULTIVO DE TEJIDOS; CULTURE D'EMBRYON; CULTURE DE TISSU; CULTURE MEDIA; DESARROLLO EMBRIONARIO; DEVELOPPEMENT EMBRYONNAIRE; DIVISION CELLULAIRE; DIVISION CELULAR; EMBRYO CULTURE; embryo development; Embryonic and Fetal Development; EMBRYONIC DEVELOPMENT; Epithelial Cells; EXPERIMENTACION IN VITRO; EXPERIMENTATION IN VITRO; FECONDATION; FECUNDACION; Female; FERTILIZATION; Fertilization in Vitro - veterinary; follicular fluid; Follicular Fluid - physiology; GANADO BOVINO; IN VITRO EXPERIMENTATION; in vitro fertilization; LIQUIDE BIOLOGIQUE; LIQUIDOS CORPORALES; Male; MEDIO DE CULTIVO; MILIEU DE CULTURE; Oocytes - growth & development; Oocytes - physiology; OVA; Ovary - physiology; OVULE; OVULO; Spermatozoa - physiology; TISSUE CULTURE
• ISSN: 0093-691X; 1879-3231
• DOI: 10.1016/S0093-691X(98)00059-4

This study was conducted to evaluate the effect of bovine follicular fluid (BFF) on fertilizability and developmental capacity of bovine oocytes matured in vitro. Oocytes were collected from slaughterhouse ovaries, and matured in TCM 199 supplemented with 5% superovulated cow serum (SCS), 2 mM pyruvate and 1 IU/mL PMSG. BFF was aspirated from small follicles (1 to 5mm in diameter). In Experiment 1, BFF was added to the Brackett and Oliphant (BO) fertilization medium at concentrations of 0, 1, 5, 10 and 20%. After insemination with frozen-thawed and heparin-treated (10 μ/mL. 15 min) bull spermatozoa for 18 h, some of the oocytes were fixed and stained to evaluate the fertilization rate. The rest of the oocytes were co-cultured in serum-free embryo culture medium (ECM; TCM199 supplemented with 5% SCS, 2 mM pyruvate and 5 μ/mL insulin) with bovine oviductal epithelial cells (BOEC) at 38.5°C under 5% CO 2 in air, and the developmental capacity of embryos was examined at 2, 7 and 9 d. In Experiment 2, BFF was added to the serum-free ECM with BOEC at 0, 5, 10 and 20% concentrations, and embryos were cultured for 9 d. Fertilization rates and blastocyst rates in low (1 and 5%) BFF in fertilization medium were not significantly different from the control (without BFF). However, high concentrations of BFF (10 and 20%) in the fertilization medium suppressed both fertilization rates and development. Large vesicles with fast monolayer formation were observed at all concentrations of BFF added to ECM with BOEC. There were no significant differences in cleavage or development to blastocyst in different concentrations of BFF added to ECM. However, the rate of development to hatched blastocysts in 20% BFF was significantly lower than that of the control (P