牛分离精子对体外受精的影响

Effect of flow cytometrically-sorted sperm on in vitro fertilization of bovine oocytes

本研究的目的是测定流动细胞分离仪分离的精子对牛卵母细胞体外受精和胚胎发育的影响。共使用了4273个牛的卵母细胞和3种类型的精子(分离、染色未分离和未染色未分离)进行体外受精试验。这些精子分别来自于3头公牛,每头公牛重复试验3～5次。数据使用ANOVA程序进行统计分析。结果显示,用3种类型精子受精后的卵母细胞囊胚率无显著差异,分别为20.4%,22.62%和22.14%,但用分离精子受精后的卵裂率显著低于未分离的精子(53.66%比71.93%,P<0.05)。所测试的3头公牛的卵裂率和囊胚率在不同公牛之间有所差异,H008号公牛所产生的囊胚明显低于H010号公牛组(分别为17.4%比25.1%,P<0.05)。结果证明分离过程或染色并没有影响胚胎发育,流动细胞分离仪可以有效地用于牛胚胎的体外生产。此外,分离、染色未分离和未分离精子的受精和胚胎发育率,在不同公牛之间有差异的这一发现表明,选择能产生高质量精子的公牛精液进行精子分离,可以提高体外受精的效率。

The objective of this study was to determine the effect of flow cytometrically-sorted, stained and unsorted sperm on in vitro fertilization and blastocyst development of bovine oocytes A total of 4273 bovine oocytes and three sperm treatments (sorted, stained and unsorted) from 3 bulls (3 to 5 replicates of each bull) were examined Data were analyzed by ANOVA procedures Analysis revealed that there was no significant difference in blastocyst rate among the three types of sperm (204% vs 2262% and 2214% ); however, the cleavage rate of oocytes fertilized with sorted sperm (5366%) was lower(P<005)than that of unsorted sperm (7193%) There were significant differences in cleavage or blastocyst rates among bulls tested H008 produced fewer(P<005) embryos than H010(174% vs 251% )The results indicated that sorting procedure or stain did not affect embryo development to blastocyst, which demonstrated that flow cytometrically-sorted sperm can be used for efficient in vitro production The finding that fertilization and embryo development rates with sorted, stained unsorted sperm were variable among bulls indicates that choosing high quality sperm from bulls for sperm sorting can increase the efficient in vitro fertilization