利用亮甲酚蓝(BCB)试验筛选基于卵泡大小的牛卵母细胞的发育能力。

Hamed Karami Shabankareh, Golshan Azimi, Mehran Torki
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摘要

背景:许多研究报道卵泡大小对卵母细胞的发育潜力起着至关重要的作用。此外,亮甲酰蓝(BCB)试验是选择更有能力的卵母细胞的最重要标准之一。目的:筛选具有发育能力的牛卵母细胞。材料与方法:采用分离卵泡的方法,从卵巢中提取牛积云卵母细胞复合物(COCs) 1730个,按卵泡直径大小(小、6 mm)分为3类。卵母细胞暴露于BCB染色,用Dulbecco的磷酸盐缓冲盐水稀释,用0.4%牛血清白蛋白(BSA)修饰90分钟。细胞质呈蓝色或不呈蓝色的卵母细胞分别被标记为BCB(+)和BCB(-)。结果:来自大卵泡和中等卵泡的BCB(+)卵母细胞和对照卵母细胞表现出较高的(p)水平。结论:目前的研究结果证实,并非每个BCB(+)卵母细胞都能导致完美的胚胎发育,BCB试验不足以鉴定能够体外胚胎发育的卵母细胞。
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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Developmental competence of bovine oocytes selected based on follicle size and using the brilliant cresyl blue (BCB) test.

Background: Many studies reported that follicle size has an essential role in developmental potential of oocytes. Also, the brilliant cresyl blue (BCB) test is one of the most important criteria in selection of more competent oocytes.

Objective: Selection of developmentally competent bovine oocytes.

Materials and methods: A total of 1730 bovine cumulus oocyte complexes (COCs) were recovered from the ovaries by follicles isolation and classified into 3 categories according to the diameters of the follicles (small, <3 mm; medium 3-6 mm and large >6 mm). Oocytes were exposed to the BCB stain, diluted in Dulbecco's phosphate-buffered saline, modified with 0.4% bovine serum albumin (BSA) for 90 min. Oocytes with or without blue coloration of the cytoplasm were designated as BCB(+) and BCB(-), respectively.

Results: The BCB(+) and control oocytes originated from large and medium follicles exhibited a higher (p<0.0001) cleavage and blastocyst rate than BCB(-) oocytes. Furthermore, the BCB(+) oocytes from large and medium follicles had the highest (p<0.0001) proportion of blastocyst than other treatment groups. In contrast, the BCB(-) oocytes from small follicles had the lowest (p<0.0001) proportion of blastocyst than other treatment groups. Interestingly, the percentage of the BCB(+) oocytes from the large and medium ovarian follicles was significantly higher (p<0.0001), than the BCB(+) oocytes from the small follicles.

Conclusion: Current results confirmed that each BCB(+) oocyte could not lead to perfect embryo development and the BCB test is not sufficient enough for the identification of oocytes that are competent for in vitro embryo development.

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