Meiotic competence of canine oocytes embedded in collagen gel.
The present study was conducted to examine the meiotic competence of canine oocytes embedded in collagen gel, and to investigate the effects of timed exposure of the oocytes embedded in collagen gel to gonadotrophins during maturation culture, on their nuclear maturation. Cumulus-oocyte complexes (COCs) were collected from bitches at the anoestrous and dioestrous stages of the reproductive cycle. In the first experiment, half of the COCs were embedded in collagen gels. The COCs with or without collagen-gel embedding were cultured in a TCM-199 medium supplemented with 0.1 IU/ml human menopausal gonadotropin (hMG) and 10 IU/ml human chorionic gonadotropin (hCG) for 72 h. In the second experiment, the COCs embedded in collagen gels were cultured in TCM-199 medium with gonadotrophins (hMG and hCG) for various periods (0, 24, 48 and 72 h) and then cultured in the medium without gonadotrophins until reaching total culture period (72 h). The percentage of the oocytes reaching metaphase I and metaphase II (MI/MII) was significantly higher (p < 0.05) in COCs with collagen-gel embedding than in COCs without collagen-gel embedding. The percentage of oocytes that were arrested at the germinal vesicle stage was significantly lower (p < 0.05) in oocytes cultured with gonadotrophins than in oocytes cultured without gonadotrophins. However, there were no significant differences in the percentages of oocytes that reached each stage of meiosis among the groups, irrespective of the duration of exposure to gonadotrophins. These observations indicate that embedding of COCs by collagen gel enhances the meiotic competence of canine oocytes, but removal of hormone supplement from maturation medium does not improve the ability of the oocytes to reach MII stage.
Otoi
T
T
Laboratory of Animal Reproduction, Department of Veterinary Science, Yamaguchi University, Yoshida, Yamaguchi, Japan. otoi@yamaguchi-u.ac.jp
Shimizu
R
R
Naoi
H
H
Wongsrikeao
P
P
Agung
B
B
Taniguchi
M
M
D000780:Anestrus / Q000502:physiology; D000818:Animals; D018929:Cell Culture Techniques / Q000662:veterinary; D003094:Collagen; D004031:Diestrus / Q000502:physiology; D004285:Dogs / Q000502:physiology*; D005260:Female; D006062:Gonadotropins / Q000494:pharmacology*; D008540:Meiosis / Q000187:drug effects* / Q000502:physiology*; D009865:Oocytes / Q000502:physiology*
D006062:Gonadotropins; D003094:Collagen