Effect of centrifugation treatment before vitrification on the viability of porcine mature oocytes and zygotes produced in vitro.
The aim of the present study was to investigate the effects of centrifugation pretreatment on the viability and nuclear status of porcine in vitro matured (IVM) oocytes and on the developmental competence of in vitro fertilized (IVF) oocytes (zygotes) after cryopreservation by vitrification (Solid Surface Vitrification; SSV). Mature oocytes having the first polar body after IVM and zygotes having the second polar body at 10 h after IVF were centrifuged at 10,000 x g at 37 C for 20 min and then subjected to SSV. Their viability was evaluated by morphological appearance and fluorescein diacetate staining. The nuclear status of oocytes was evaluated 6 h after vitrification. The developmental ability to the blastocyst stage of vitrified zygotes was evaluated after 6 days of in vitro culture. Although centrifugation did not damage the oocytes directly, it drastically reduced the rate of live oocytes after SSV. The rates of vitrification-induced parthenogenetic activation were similar in both centrifuged and non-centrifuged oocytes (42.4 and 47.4%, respectively). Centrifugation had no significant effects on the viability of pronuclear oocytes. The development of vitrified zygotes to the blastocyst stage was significantly lower than that of the control irrespective of centrifugation pretreatment. There was no difference in the cleavage and blastocyst rates between the control and centrifuged zygotes after vitrification. There was also no difference in the total cell numbers of blastocysts between the control and centrifuged zygotes irrespective of vitrification. These results reveal that, in IVM porcine oocytes, centrifugation pretreatment is highly detrimental to cryotolerance; however, in zygotes, it has only a slight effect on viability and does not alter the developmental competence of surviving zygotes.
Somfai
Tamás
T
Division of Animal Sciences, National Institute of Agrobiological Sciences, Tsukuba, Japan.
Kashiwazaki
Naomi
N
Ozawa
Manabu
M
Nakai
Michiko
M
Maedomari
Naoki
N
Noguchi
Junko
J
Kaneko
Hiroyuki
H
Nagai
Takashi
T
Kikuchi
Kazuhiro
K
D000818:Animals; D002470:Cell Survival; D002498:Centrifugation; D015925:Cryopreservation; D005260:Female; D005307:Fertilization in Vitro / Q000379:methods* / Q000662:veterinary*; D066298:In Vitro Techniques; D009865:Oocytes / Q000166:cytology*; D034421:Sus scrofa; D015053:Zygote / Q000166:cytology*