Effect of post-activation culture conditions on the development of parthenogenetic embryos in cattle

A. Lopukhov
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Abstract

Purpose: to conduct a comparative analysis of the effect of commercial media BO-IVC and СR1aa at the stage of the activation and subsequent culture of artificially activated oocytes on the formation and quality of parthenogenetic bovine embryos.Materials and methods. 3 groups of disemeters of 50 goals in each were formed. In the first experimental group, the disemeted was in a meticulous manner with a ram-industrialist (artificial kriproporchid), in the second experimental-with a penEexctomed ram-industrialist. In the third (control) group, a producer ram was used. In the first experimental group of a ram-industrialist (artificial kriproporchid) with attached taps were released into a group of sheep twice a day for 1.5-2 hours. In the second experimental group of a penEctomed ram, it was placed in the corral to the disemetery in the morning for 3 hours. In the third group, the lamb producer was constantly with the disemets for two weeks, then he was changed on a new ram i.e. Used the methodology used in the farm. During the experiment, they observed the behavior of animals of all groups. In the experimental groups, after the detection of disemeters in the hunt, their natural insemination of the manufacturer was carried out. Based on the results of the subsequent oster, the effectiveness of the reproduction of sheep was evaluated.Results. The cleavage rate did not differ between the experimental groups, varying from 73,0 to 76,5%. Also, there was not found a significant effect of the conditions for post-activation culture of oocytes on their development before late morula and late blastocyst stage, which was for the CR1aa/CR1aa, CR1aa/BO-IVC and BOIVC/ BO-IVC groups 28,9±1,7, 40,4±7,5 and 36,0±6.4%, respectively. Meanwhile, we found out the effect of tested culture conditions on the ability of parthenogenetic embryos to overcome the 8-16 cell block and their quality on the late stages of embryo development. The rate of embryos with less than 16 nuclei was the highest in the CR1aa/CR1aa group (56,8±2,1 %). The replacement of CR1aa medium to BO-IVC medium (BO-IVC/BO-IVC group) significantly reduced this level (p<0,05). The positive effect was enhanced when CR1aa medium was used at the stage of culture in the presence of 6-DMAP and cycloheximide, and subsequent embryo development was in BO-IVC medium (CR1aa/BO-IVC group) (p<0.001). Furthermore, when we used the mixed variant of culture, the total cell number in parthenogenetic morula and blastocyst stage embryos increased (p<0.05).Conclusion. Thus, the BO-IVC medium at the stages of post-activation and subsequent development of artificially activated bovine oocytes is comparable to the CR1aa medium in terms of the efficiency of obtaining parthenogenetic embryos at the blastocyst stage. Nevertheless, its replacement at the post-activation stage with CR1aa medium makes it possible to improve the quality of parthenogenetic embryos.
活化后培养条件对牛孤雌胚胎发育的影响
目的:比较分析人工激活卵母细胞激活及后续培养阶段,商业培养基BO-IVC和СR1aa对孤雌牛胚胎形成及质量的影响。材料和方法。分为3组,每组50个目标。在第一个实验组中,被试者以一种细致的方式与一个公羊工业家(人工kriproporchid)相处,在第二个实验中,与一个阴茎切除的公羊工业家相处。在第三组(对照组)中,使用生产者公羊。在第一组实验中,将一个带有水龙头的公羊-工业家(人造kriproporchid)每天两次释放到一组绵羊中,持续1.5-2小时。第二组实验组为一只阴茎切除公羊,于早晨将公羊置于离墓地的畜栏中3小时。在第三组中,羔羊饲养员连续两周与猪在一起,然后他被换了一只新的公羊,即使用农场使用的方法。在实验过程中,他们观察了各组动物的行为。实验组在狩猎中检测到异径蝶后,对其进行制造者自然授精。在后续试验结果的基础上,对绵羊的繁殖效果进行了评价。不同实验组的卵裂率差异不大,在73.3% ~ 76.5%之间。CR1aa/CR1aa组、CR1aa/BO-IVC组和BOIVC/ BO-IVC组卵母细胞在晚黑子期和囊胚期前发育的差异无统计学意义(分别为28.9±1,7,40,4±7,5和36.0±6.4%)。同时,我们还发现了不同的培养条件对孤雌生殖胚胎克服8-16细胞阻滞的能力的影响及其对胚胎发育后期质量的影响。CR1aa/CR1aa组胚胎少于16个细胞核的比例最高(56.8±2.1%)。将CR1aa培养基替换为BO-IVC培养基(BO-IVC/BO-IVC组)可显著降低该水平(p< 0.05)。在6-DMAP和环己亚胺的作用下,在培养阶段使用CR1aa培养基增强了这种积极作用,随后胚胎发育在BO-IVC培养基中(CR1aa/BO-IVC组)(p<0.001)。此外,当我们使用混合变体培养时,孤雌生殖桑葚胚和囊胚期胚胎的细胞总数增加(p<0.05)。因此,在人工激活牛卵母细胞后激活和随后发育阶段的BO-IVC培养基在囊胚期获得孤雌胚胎的效率方面与CR1aa培养基相当。然而,在激活后阶段用CR1aa培养基替代它可以提高孤雌胚胎的质量。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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