Developmental competence of in vitro-fertilized porcine oocytes after in vitro maturation and solid surface vitrification: effect of cryopreservation on oocyte antioxidative system and cell cycle stage.
The susceptibility of in vitro matured (IVM) porcine oocytes to be fertilized in vitro after vitrification was investigated. IVM oocytes were cryopreserved by solid surface vitrification (SSV) or treated with cryoprotectants (toxicity control, TC). Control oocytes were not treated or vitrified. Live oocytes in the three groups were in vitro fertilized (IVF) and then cultured (IVC) for 6 days. In vitro maturation and IVC were performed under 5% or 20% O(2) tension. The percentage of live oocytes in the SSV group was lower than those in the control and TC groups. Fertilization rates after SSV were significantly lower than in the control group. Significantly fewer penetrated oocytes formed male pronuclei in the SSV group than in the control and TC groups. Cleavage rates were significantly lower in the SSV group than in the control and TC groups. Blastocyst formation rates in the control and TC groups were similar, whereas only a single embryo developed to the blastocyst stage from 113 oocytes after vitrification. Blastocyst formation rates in the control group and in the TC group were significantly higher under 5% O(2) IVC than under 20% O(2) IVC. Oxygen tension during IVM had no effect on embryo development. The glutathione (GSH) content of vitrified oocytes was significantly lower than in the controls. In contrast, the H(2)O(2) level was higher in vitrified oocytes than in control oocytes. Vitrification caused parthenogenetic activation in 44.9% of unfertilized oocytes. This significant increase in parthenogenetic activation along with significantly dropped GSH level in vitrified oocytes may explain the decreased ability of the SSV group to form male pronuclei. These factors might have contributed to the poor developmental competence of vitrified oocytes.
Genetic Diversity Department, National Institute of Agrobiological Sciences, Kannondai 2-1-2, Tsukuba, Ibaraki, Japan.
Tamás
T
Somfai
Manabu
M
Ozawa
Junko
J
Noguchi
Hiroyuki
H
Kaneko
Ni Wayan
NW
Kuriani Karja
Mokhamad
M
Farhudin
András
A
Dinnyés
Takashi
T
Nagai
Kazuhiro
K
Kikuchi
D000818:Animals; D002453:Cell Cycle / Q000502:physiology*; D002478:Cells, Cultured; D015925:Cryopreservation; D047108:Embryonic Development / Q000502:physiology*; D005307:Fertilization in Vitro; D009865:Oocytes / Q000502:physiology*; D018384:Oxidative Stress / Q000502:physiology*; D010100:Oxygen / Q000502:physiology; D013552:Swine
D010100:Oxygen