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Cushioned centrifugation during sperm selection increases the fertilization and cleavage rates of cattle embryos produced in vitro.
Animal Reproduction Science ( IF 2.2 ) Pub Date : 2020-05-19 , DOI: 10.1016/j.anireprosci.2020.106508
Cecilia Urquiza Machado Pavin 1 , Fabio Gallas Leivas 1 , Francielli Weber Santos 1 , Daniele Missio 1 , Fernando Silveira Mesquita 1 , Daniela Dos Santos Brum 1
Affiliation  

This study was conducted to evaluate the effect of utilization of an iodixanol-based solution as a cushioning method during the sperm selection utilizing discontinuous Percoll gradient centrifugation in in vitro production (IVP) of cattle embryos. In Experiment I, all aliquots of thawed semen were subjected to sperm selection using the same discontinuous Percoll® gradients, except for the following four conditions: presence of cushioning solution (Cushion Fluid, Minitube) during the first centrifugation process (C1), presence of cushioning solution during the second centrifugation process (C2), inclusion of cushioning solution in both centrifugation steps (C1-2), and no addiction of cushioning solution (C; control group). Recovery rates, sperm kinetics, and reactive oxygen species (ROS) production were evaluated. In Experiment II, sperm cells were processed using sperm selection conditions C and C1, and fertilization rates and embryonic development kinetics were compared between experimental groups. With use of condition C1, there was improvement in fertilization and cleavage rates when compared to use of condition C (56.4% compared with 45.5% and 80.0% compared 64.7%, respectively). In conclusion, results indicate the use of a cushioning solution during sperm selection positively affects the developmental potential of embryos.



中文翻译:

精子选择过程中的缓冲离心增加了体外产生的牛胚胎的受精率和卵裂率。

进行这项研究以评估在体外使用不连续Percoll梯度离心法在选择精子过程中使用碘克沙醇溶液作为缓冲方法的效果牛胚胎的生产(IVP)。在实验I中,除以下四个条件外,所有解冻后的精液等分试样均使用相同的不连续Percoll®梯度进行精子选择:第一次离心过程(C1)中存在缓冲液(Cushion Fluid,Minitube),在第二次离心过程(C2)中添加缓冲液,在两个离心步骤(C1-2)中都包含缓冲液,并且缓冲液没有上瘾(C;对照组)。评估回收率,精子动力学和活性氧(ROS)产生。在实验II中,使用精子选择条件C和C1处理精子细胞,并比较实验组之间的受精率和胚胎发育动力学。使用条件C1,与使用条件C相比,受精率和卵裂率都有改善(分别为56.4%和45.5%,80.0%和64.7%)。总之,结果表明在选择精子过程中使用缓冲液会积极影响胚胎的发育潜力。

更新日期:2020-06-25
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