Yashiro Ikuko, Tagiri Miho, Ogawa Hayato, Tashima Kazuya, Takashima Seiji, Hara Hiromasa, Hirabayashi Masumi, Hochi Shinichi
Graduate School of Science and Technology Faculty of Textile Science and Technology Shinshu University, Nagano 386-8567, Japan National Institute for Physiological Sciences Aichi 444-8787, Japan.
Graduate School of Science and Technology Faculty of Textile Science and Technology Shinshu University, Nagano 386-8567, Japan National Institute for Physiological Sciences Aichi 444-8787, Japan Graduate School of Science and Technology Faculty of Textile Science and Technology Shinshu University, Nagano 386-8567, Japan National Institute for Physiological Sciences Aichi 444-8787, Japan.
Reproduction. 2015 Apr;149(4):347-55. doi: 10.1530/REP-14-0594. Epub 2015 Jan 27.
The objective of this study was to investigate whether developmental competence of vitrified-warmed bovine oocytes can be improved by antioxidant treatment during recovery culture. In experiment 1, one of the two antioxidants (either l-ascorbic acid or α-tocopherol) was added as a supplement to the recovery culture medium to which postwarming oocytes were exposed for 2 h before IVF. The exposure to α-tocopherol had a positive effect on rescuing the oocytes as assessed by the blastocyst yield 8 days after the IVF (35.1-36.3% vs 19.2-25.8% in untreated postwarming oocytes). Quality of expanding blastocysts harvested on Day 8 was comparable between α-tocopherol-treated vitrification group and fresh control group in terms of total cell number and chromosomal ploidy. In experiment 2, level of reactive oxygen species, mitochondrial activity, and distribution of cortical granules in α-tocopherol-treated postwarming oocytes were assessed. No obvious differences from the control data were found in these parameters. However, the treatment with α-tocopherol increased the percentage of zygotes exhibiting normal single aster formation (90.3% vs 48.0% in untreated postwarming oocytes; 10 h post-IVF). It was concluded that α-tocopherol treatment of vitrified-warmed bovine mature oocytes during recovery culture can improve their revivability, as shown by the high blastocyst yield and the higher mean total cell number in the blastocysts.
本研究的目的是调查在复苏培养过程中进行抗氧化剂处理是否可以提高玻璃化冷冻-解冻牛卵母细胞的发育能力。在实验1中,两种抗氧化剂之一(l-抗坏血酸或α-生育酚)作为补充剂添加到复苏培养基中,解冻后的卵母细胞在体外受精前在该培养基中暴露2小时。通过体外受精后8天的囊胚产量评估,暴露于α-生育酚对挽救卵母细胞有积极作用(35.1-36.3% 对比未处理的解冻后卵母细胞的19.2-25.8%)。在总细胞数和染色体倍性方面,在第8天收获的扩张囊胚的质量在α-生育酚处理的玻璃化冷冻组和新鲜对照组之间具有可比性。在实验2中,评估了α-生育酚处理的解冻后卵母细胞中的活性氧水平、线粒体活性和皮质颗粒的分布。在这些参数中未发现与对照数据有明显差异。然而,α-生育酚处理增加了表现出正常单星体形成的受精卵的百分比(90.3% 对比未处理的解冻后卵母细胞的48.0%;体外受精后10小时)。得出的结论是,在复苏培养过程中用α-生育酚处理玻璃化冷冻-解冻的牛成熟卵母细胞可以提高其复苏能力,如高囊胚产量和囊胚中更高的平均总细胞数所示。