Missio Daniele, Folchini Natália Picolli, Leivas Fabio Gallas, Pavin Cecília Isabel Inês Urquiza Machado, Pinto Hirya Fernandes, Cibin Francielli Weber Santos, Brum Daniela Dos Santos
Federal University of Pampa (UNIPAMPA), BIOTECH, Laboratory Biotechnology of Reproduction, 97.500-970,Uruguaiana, RS, Brazil.
Federal University of Pampa (UNIPAMPA), BIOTECH, Laboratory Biotechnology of Reproduction, 97.500-970,Uruguaiana, RS, Brazil.
Anim Reprod Sci. 2018 May;192:146-153. doi: 10.1016/j.anireprosci.2018.03.002. Epub 2018 Mar 4.
The aim of the present study was to evaluate the effects of Percoll volume on recovery rate, sperm quality, and embryo development kinetics in in vitro production of cattle embryos. Straws of conventional and sex-sorted semen were allocated to three different volumes of Percoll: 300 μL of each Percoll gradient (90%, 60%, and 30%), Control; 100 μL of each Percoll gradient, P100; and 200 μL of each Percoll gradient, P200. Sperm quality, fertilization rate, and embryo morpho-kinetic development using time lapse cinematography up to 48 h post-insemination were evaluated. For conventionally processed semen, sperm motility, vigor, and recovery rate were greater in the P100 and P200 treatment groups compared to the Control (P < 0.05), whereas reactive oxygen species (ROS) concentrations, lipid peroxidation, and superoxide dismutase enzyme activity were not influenced by treatments. For sex-sorted semen, treatment with P100 increased sperm curvilinear velocity, average path velocity, and amplitude of lateral head displacement (P < 0.05). Recovery rate was greater in the P100 group than Control and P200 groups (P < 0.05), formation of ROS was less in the P100 than Control and P200 groups, and superoxide dismutase enzyme activity was less in the P100 than Control group. Fertilization and cleavage rates, time of first cleavage, and cell number were similar between the P100 and Control groups (P > 0.05). The inclusion of Percoll volumes of 100 μL resulted in an increased sperm recovery rate without damage to sperm quality or affecting early embryonic development.
本研究的目的是评估在体外生产牛胚胎过程中，Percoll体积对回收率、精子质量和胚胎发育动力学的影响。将常规精液和性别分选精液的细管分配到三种不同体积的Percoll中：每种Percoll梯度（90%、60%和30%）300μL，对照组；每种Percoll梯度100μL，P100组；每种Percoll梯度200μL，P200组。评估了精子质量、受精率以及使用延时摄影技术在授精后48小时内观察到的胚胎形态动力学发育情况。对于常规处理的精液，与对照组相比，P100和P200处理组的精子活力、活率和回收率更高（P<0.05），而活性氧（ROS）浓度、脂质过氧化和超氧化物歧化酶活性不受处理的影响。对于性别分选精液，P100处理可提高精子曲线速度、平均路径速度和头部侧摆幅度（P<0.05）。P100组的回收率高于对照组和P200组（P<0.05），P100组产生的ROS比对照组和P200组少，P100组的超氧化物歧化酶活性比对照组低。P100组和对照组之间的受精率、卵裂率、首次卵裂时间和细胞数量相似（P>0.05）。加入100μL 的Percoll可提高精子回收率，且不会损害精子质量或影响早期胚胎发育。
