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
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
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DOI：
10.1016/j.cryobiol.2007.06.008
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发表时间：
2007-10-01
期刊：
影响因子：
2.7
通讯作者：
Kikuchi, Kazuhiro
中科院分区：
文献类型：
--
作者：
Tamas Somfai;Ozawa, Manabu;Kikuchi, Kazuhiro
研究了体外成熟（IVM）猪卵母细胞玻璃化冷冻后体外受精的敏感性。 IVM 卵母细胞通过固体表面玻璃化 (SSV) 冷冻保存或用冷冻保护剂（毒性控制，TC）处理。对照卵母细胞未经处理或玻璃化。三组活卵母细胞均进行体外受精（IVF），然后培养（IVC）6天。体外成熟和IVC在5％或20％O 2 张力下进行。 SSV组的活卵母细胞百分比低于对照组和TC组。 SSV 后的受精率显着低于对照组。 SSV 组中形成雄性原核的穿透卵母细胞明显少于对照组和 TC 组。 SSV 组的卵裂率显着低于对照组和 TC 组。对照组和 TC 组的囊胚形成率相似，但玻璃化冷冻后 113 个卵母细胞中只有一个胚胎发育到囊胚阶段。 5% 02 IVC 下对照组和 TC 组的囊胚形成率显着高于 20% O-2 IVC 下。 IVM 期间的氧张力对胚胎发育没有影响。玻璃化卵母细胞的谷胱甘肽（GSH）含量显着低于对照。相反，玻璃化卵母细胞中的 H2O2 水平高于对照卵母细胞。玻璃化冷冻导致 44.9% 的未受精卵母细胞孤雌生殖激活。这种孤雌生殖激活的显着增加以及玻璃化卵母细胞中 GSH 水平的显着下降可能解释了 SSV 组形成雄性原核的能力下降。这些因素可能导致玻璃化卵母细胞发育能力差。 (c) 2007 Elsevier Inc. 保留所有权利。
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% 02 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% 02 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 H2O2 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. (c) 2007 Elsevier Inc. All rights reserved.