亮甲酚蓝染色对牛卵母细胞体外成熟的影响

Effect of Brilliant Cresyl Blue on in Vitro Maturity of Bovine Oocytes

研究旨在观察亮甲酚蓝(brilliant cresyl blue,BCB)对牛卵母细胞的染色效果及体外成熟的影响,从而筛选出更高质量的卵母细胞用于胚胎工程研究。从本地屠宰场收集牛卵巢后采用抽吸法获取卵泡液,在体视显微镜下选择卵丘完整、胞质均匀的卵丘-卵母细胞复合体(COCs)随机分组,分别进行BCB染色时间和不同浓度BCB染色的试验。结果表明,染色90 min后卵母细胞着色率为68.1%,显著高于30、60 min组(P<0.05),辨识难度为"中";39μmol/L组着色率显著高于13μmol/L组(P<0.05),与26、52μmol/L组差异不显著(P>0.05),成熟数最高(62),成熟率显著高于52μmol/L组,与其它组差异不显著(P>0.05);BCB+组和BCB-组的成熟率与对照组比较差异均不显著(P>0.05)。结果提示,39μmol/L BCB染色90 min能有效筛选牛COCs用于IVM,但是并不能显著提高牛卵母细胞的成熟率。

This experiment aimed at the observation of the staining effect of brilliant cresyl blue（ BCB）and its influence on bovine oocyte in vitro maturation so as to select the higher quality oocytes for the embryo engineering research. Bovine ovaries were collected from a local slaughterhouse,and with the suction method the follicular fluid was obtained,cumulus oocyte complexes（ COCs） with intact cumulus,homogeneous cytoplasm were selected by using a stereoscopic microscope,and then were randomly divided into two groups,and the staining time and BCB concentration were determined,respectively. The results showed that,the staining rate of oocyte 90 min after staining was 68. 1%,it was significantly higher than that of the groups of 30 and 60 min,and the distinguishable difficulty was＂common＂. The rate of stained oocytes in the 39 μmol /L group was significantly higher than that in the 13 μmol /L group（ P 〈0.05）,and was not different from that in the 26 and52 μmol /L groups（ P 〉0.05）,the No. of mature oocytes was the highest（ 62）,the maturation rate was significantly higher than that in the 52 μmol /L group（ P 〉0.05）,and was not different from that in the other groups（ P 〈0.05）. The maturation rates of the BCB＋group and BCB-group were not significantly different from that of the control group（ P 〉0.05）. These results suggest that,staining with 39 μmol /L of BCB for 90 min is effective for selection of bovine COCs for IVM,but has no significant efficiency in improving the rate of bovine oocyte in vitro maturation.