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Effects of bovine spermatozoa preparation on embryonic development in vitro

机译:牛精子制剂对体外胚胎发育的影响

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摘要

The aim of our research was to examine the ability of density gradient preparation BoviPure® and swim up method on bull sperm separation and in vitro embryo production (IVP) systems. Frozen/thawed semen from six Simmental bulls was pooled and treated using both methods. The sperm motility, concentration, membrane activity, membrane integrity and acrosomal status were evaluated and compared before and after sperm processing using BoviPure® and swim up methods. We also evaluated and compared cleavage rates, embryo yield and quality between the methods. There were significant differences (P < 0.05) between the sperm characteristics before and after BoviPure®, but not after swim up method. However, there were significant differences for sperm results among those two mentioned methods. A total of 641 oocytes were matured and fertilized in vitro and cultured in SOFaaBSA. The percentage of cleavage (Day 2) and the percentage of hatched embryos (Day 9) were similar for both methods. However, embryo production rate (Day 7) was significantly higher using BoviPure® method (P < 0.05). Also, total cell number and embryo differential staining (inner cell mass and trophectoderm cells) of Day 7 morulas and blastocysts showed that BoviPure® treated sperm displayed higher quality embryos compared to swim up method (P < 0.05). Our results indicate that BoviPure® method has an enhanced capacity in sperm selection for in vitro embryo production when compared with swim up method. So, we concluded that BoviPure® could be considered as a better alternative to swim up method for separating bull spermatozoa from frozen/thawed semen for IVP of bovine embryos.
机译:我们的研究目的是检验密度梯度制备BoviPure ®的能力和游动方法对公牛精子分离和体外胚胎产生(IVP)系统的影响。合并来自六只西门塔尔牛的冷冻/解冻精液,并使用两种方法进行处理。使用BoviPure ®和游泳方法评估和比较精子处理前后的精子运动性,浓度,膜活性,膜完整性和顶体状态。我们还评估和比较了两种方法之间的卵裂率,胚胎产量和质量。 BoviPure ®之前和之后的精子特征之间存在显着差异(P <0.05),但在游泳方法后没有。但是,在上述两种方法之间,精子结果存在显着差异。总共641个卵母细胞在体外成熟并受精并在SOFaaBSA中培养。两种方法的卵裂百分比(第2天)和孵化胚胎的百分比(第9天)相似。但是,使用BoviPure ®方法时,胚胎的生产率(第7天)明显更高(P <0.05)。另外,第7天桑和胚泡的总细胞数和胚胎分化染色(内部细胞团和滋养外胚层细胞)显示,与游泳法相比,经BoviPure ®处理的精子显示出更高质量的胚胎(P <0.05) 。我们的结果表明,与游泳法相比,BoviPure ®方法具有更高的精子选择能力,可用于体外胚胎生产。因此,我们得出结论,可以认为BoviPure ®可以作为游走方法的更好替代方法,用于从牛胚胎的IVP中分离冷冻/解冻精液中的公牛精子。

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