Effects of ovary storage time and temperature on DNA fragmentation and development of porcine oocytes.
The present study was conducted to investigate the effects of storage time and temperature of porcine ovaries on the quality and nuclear maturation in vitro of oocytes obtained from stored ovaries and their subsequent development after in vitro fertilization. The ovaries were stored in physiological saline for 0, 3, 6, 9 and 12 h at various temperatures (4, 15, 25 and 35 C). The pH of follicular fluid obtained from the ovaries, DNA fragmentation of the oocyte nucleus and meiotic competence of oocytes were examined. Some oocytes from ovaries stored at 15, 25 and 35 C for 6 h were fertilized in vitro, and then cultured for 7 days to examine the ability of embryos to develop to the blastocyst stage. When the ovaries were stored at 35 C, the pH of follicular fluid decreased and the proportions of oocytes with DNA fragmented nuclei increased as the storage time was prolonged, and the storage of ovaries for 6, 9 and 12 h resulted in lower maturation rates of oocytes. When the ovaries were stored at 4, 15, 25 and 35 C for 6 h, the storage at higher temperatures (> or =15 C) decreased the pH of follicular fluid and induced nucleic DNA fragmentation in higher proportions of oocytes. None of the oocytes from ovaries stored at 4 C reached metaphase II. The storage of ovaries at 15 C reduced the rates of in vitro fertilized oocytes and subsequent embryo development, but there were no significant differences in the rates of fertilization and blastocyst formation between oocytes from ovaries stored at 25 C and 35 C. Our findings indicate that the storage of ovaries at 25-35 C for 6 h is effective for maintaining the developmental competence of porcine oocytes even though the development rates were lower than those of ovaries stored at 35 C for 3 h.
Wongsrikeao
Pimprapar
P
Laboratory of Animal Reproduction, The United Graduate School of Veterinary Science, Yamaguchi University, Japan.
Otoi
Takeshige
T
Karja
Ni Wayan Kurniani
NW
Agung
Budiyanto
B
Nii
Masahiro
M
Nagai
Takashi
T
D000818:Animals; D001755:Blastocyst / Q000378:metabolism; D002467:Cell Nucleus / Q000378:metabolism; D046508:Culture Techniques; D004249:DNA Damage; D053938:DNA Fragmentation; D047108:Embryonic Development; D005260:Female; D005307:Fertilization in Vitro; D006863:Hydrogen-Ion Concentration; D020287:In Situ Nick-End Labeling; D008297:Male; D009865:Oocytes / Q000378:metabolism*; D006080:Ovarian Follicle / Q000378:metabolism; D010053:Ovary / Q000378:metabolism* / Q000473:pathology; D013048:Specimen Handling / Q000379:methods*; D013094:Spermatozoa / Q000378:metabolism; D013552:Swine; D013696:Temperature; D013997:Time Factors