The purpose of this study was to investigate the effects of the collection time, co-culture and sperm penetration of canine oocytes on in vitro maturation and fertilization. The oocytes were cultured in TCM-199 media containing hormonal supplements (10% FCS, 10 IU/ml HCG, 10 IU/ml PMSG) at 5% CO2, 95% air, 38℃. The in vitro maturation rate to MⅡ stage of in vitro oocytes recovered from ovaries that collected at follicular, luteal and inactive phases of the reproductive phase for 44～72 hrs were 19.2%, 12.2%, and 6.0%, respectively. Follicular phases oocytes had a significantly higher in vitro maturation rate than oocytes collected at luteal and anestrus stage (p<0.05). The in vitro maturation rates to the MII stage of canine oocytes after 48 hrs of culture with glutathione, pyruvate, or glutathione + pyruvate were 12.5%, 10.7%, and 17.5%, respectively. This was higher than that in both alone or the combination of the two compared to the control group (19.0%). The sperm penetration rates of in vitro matured oocytes by fresh and frozen semen were 29/80 (36.3%) and 18/80 (22.5%), respectively. Although there are limited reports about canine oocytes co-culture and in vitro fertilization, our results on in vitro maturation is comparable to the results from other researches.

ABSTRACT   INTRODUCTION   MATERIALS AND METHODS   Recovery of Oocytes    In Vitro Maturation of Oocytes    Assessment of Meiotic Stage    Statistical Analysis   RESULTS    Nuclear Developmental rate of Oocytes    IVM Rates of Oocytes Co-cultured with Glutathioneand Pyruvate    Penetration Rate of Oocytes by Fresh and FrozenSperm   DISCUSSION   REFERENCES

• 박지훈(충남대학교) | Ji-Hoon Park
• Ho-Bong Seok(College of Bio-resources Science, Dankook University)
• Sang Keun Kim(College of Veterinary Medicine, Chungnam National University) Corresponding author