亮甲酚蓝优选哺乳动物卵母细胞技术研究进展
2016-10-20王俊敏唐易张锦
王俊敏 唐易 张锦
摘要 随着动物胚胎工程技术的不断发展,卵母细胞的需求量不断增大。卵母细胞的质量决定植入前早期胚胎发育的效率,为此筛选出优质卵母细胞对于提高后续胚胎的发育起着决定性的作用。本文就亮甲酚蓝染色筛选成熟培养前的卵母细胞,对于卵母细胞质量、后续胚胎发育效率和早期胚胎发育相关基因表达的影响方面的研究予以综述,期望对于同行有一定的借鉴意义。
关键词 亮甲酚蓝;卵母细胞;胚胎发育;胞质成熟
中图分类号 S813.3 文献标识码 A 文章编号 1007-5739(2016)08-0238-01
Abstract With the continuous development of animal embryo engineering and technology,the demand of oocytes is increasing a lot.Oocyte quality can affect the efficiency of early embryonic development,so it plays a decisive role for subsequent embryonic development through selecting high-quality oocytes.In this paper,the effects of Brilliant Cresol Blue staining(BCB)on further embryo development were reviewed,which included the quality and efficiency of subsequent embryonic development and the influence of gene expression of early embryo.It was expected that it would be helpful for their peers in the early embryo development research fields.
Key words brilliant cresyl blue(BCB);oocyte;embryo development;cytoplast maturation
生长期的卵母细胞代谢旺盛,含有大量的葡萄糖-6-磷酸脱氢酶(G6PDH),但是随着卵母细胞的生长期结束其活性也逐渐降低直至消失,因此卵母细胞成熟时G6PDH的含量很少。卵母细胞经过采用亮甲酚蓝(BCB)染色,由于G6PDH可以使BCB由原来的蓝色变成无色,因此可以分辨出生长期的卵母细胞和成熟的卵母细胞,前者表现为胞质未着色(BCB-),而后者则胞质呈蓝色(BCB+)。因此,BCB染色技术已经被广泛用于多种动物卵母细胞的筛选,并取得了很好的效果[1-4]。此外,已有的研究认为,处于生长阶段的卵母细胞的发育能力远低于成熟期的卵母细胞,因此通过BCB染色技术还能够间接的评估卵母细胞的质量。通过采用BCB对成熟培养前的猪[1- 2]、山羊[3]、绵羊[5]和牛[6-7]卵母细胞染色用于检测葡萄糖-6-磷酸脱氢酶间接评价优选卵母细胞,为后续研究提供优质卵母细胞。有研究发现未着色的卵母细胞培养成熟后,用于核移植具有更高的体外囊胚发育率[8- 9]。……
