Rodríguez-González Elisabeth, López-Bejar Manel, Izquierdo Dolors, Paramio María-Teresa
Departament de Ciència Animal i dels Aliments, Facultat de Vetèrinaria, Universitat Autònoma de Barcelona, 08193 Bellaterra, Barcelona, Spain.
Reprod Nutr Dev. 2003 Mar-Apr;43(2):179-87. doi: 10.1051/rnd:2003012.
The aim of this study was to assess the effect of oocyte selection using the brilliant cresyl blue (BCB) test plus the addition of cysteamine to the in vitro maturation (IVM) medium to improve the in vitro embryo development of prepubertal goat oocytes. The oocytes were exposed to 26 microM BCB and classified according to their cytoplasm coloration: BCB+ (oocytes with blue cytoplasm) and BCB- (unstained oocytes). The oocytes were matured in a conventional IVM medium supplemented with cysteamine 100 microM. The control group consisted of oocytes not exposed to BCB and matured without cysteamine. The IVM-oocytes were inseminated and cultured in synthetic oviductal fluid (SOF) for 7 days. The normal fertilisation rate (oocytes showing 2 pronuclei and 1 sperm tail) of BCB+ oocytes (40%) was higher than those of BCB- (21%) and control oocytes (22%). The percentage of morulae plus blastocysts was higher (P < 0.05) in the BCB+ group than in the BCB- group (23.8 vs. 5.1%, respectively). In conclusion, the integration of the BCB test and the addition of cysteamine in the protocol of in vitro embryo production from prepubertal goat oocytes has improved the developmental rates of embryo development.
本研究的目的是评估使用灿烂甲酚蓝(BCB)试验进行卵母细胞选择,并在体外成熟(IVM)培养基中添加半胱胺,以改善青春期前山羊卵母细胞的体外胚胎发育情况。将卵母细胞暴露于26微摩尔的BCB中,并根据其细胞质着色情况进行分类:BCB +(细胞质呈蓝色的卵母细胞)和BCB -(未染色的卵母细胞)。卵母细胞在补充有100微摩尔半胱胺的常规IVM培养基中成熟。对照组由未暴露于BCB且在无半胱胺的情况下成熟的卵母细胞组成。将IVM卵母细胞进行授精,并在合成输卵管液(SOF)中培养7天。BCB +卵母细胞的正常受精率(显示2个原核和1条精子尾巴的卵母细胞)为40%,高于BCB -卵母细胞(21%)和对照卵母细胞(22%)。BCB +组中桑葚胚加囊胚的百分比高于BCB -组(分别为23.8%和5.1%,P < 0.05)。总之,在青春期前山羊卵母细胞的体外胚胎生产方案中整合BCB试验和添加半胱胺,提高了胚胎发育率。