亮甲酚蓝染色对水牛卵母细胞体外受精和孤雌激活胚胎发育的影响

摘要

[ Objective ] This paper aims to evaluate the effects of brilliant cresyl blue (BCB) staining on the efficiency of in vitro fertilization ( IVF)and parthenogenetic activation(PA) of oocytes in buffalo. [ Method] Cumulus oocyte complexes (COCs) are stained with BCB (26 μ mol/L) for 90 min before in vitro maturation (IVM). Subsequently, COCs are divided into BCB-group (colorless cytoplasm) and BCB +group (colored cytoplasm) according to their ooplasm BCB coloration. The COCs without BCB staining are considered as control group, and the COCs of holding control group are incubated in DPBS supplemented with 0. 4％ BSA for 90 min. After IVM, the maturation rate, embryo development rate of IVF and PA in each group are evaluated. [ Result] The maturation rate in BCB + group(65.70％ ) is significantly higher than that of control group (59.86％), holding control group (58.42％) and BCB-group (48.86％). The blastocyst rate of IVF in BCB +group (27.08％) is significantly higher than that of holding control group (20.50％) and BCB-group( 8.45％ ), but there is no significant difference between the BCB + group and the control group( 27.08％ vs 25.49％ ). There is no significant difference in parthenogenetic blastocyst rate among the BCB + group, control group and holding control group( 19.13％ ), but all of the three groups are significantly higher than that of BCB-group. [ Conclusion] The positive buffalo COCs selected by BCB staining before IVM have superior developmental competence in maturation, but it could not significantly enhance the developmental rate of the IVF and PA embryos.%[目的]探讨亮甲酚蓝(BCB)染色对水牛卵母细胞体外受精及孤雌激活胚胎发育的影响.[方法]卵丘卵母细胞复合体(COCs)在体外成熟培养前用浓度26 μmol/L的亮甲酚蓝染色90 min,然后根据卵母细胞胞质蓝色的有无分为阳性(BCB+,蓝色)和阴性组(BCB-,不着色).以未经染色处理的COCs为对照组,控制对照组卵母细胞在含0.4%牛血清白蛋白(BSA)的杜氏磷酸盐缓冲液(DPBS)中孵育90 min.体外成熟培养后统计各组的成熟率、体外受精和孤雌激活后的发育率.[结果]BCB+组的成熟率(65.70%)显著高于对照组(59.86%)、控制对照组(58.42%)和BCB-组(48.86%).在体外受精后的囊胚发育率方面,BCB+组(27.08%)和对照组(25.49%)差异不显著,但显著高于控制对照组(20.50%)和BCB-组(8.45%).在孤雌激活后的囊胚发育率方面,BCB-组显著低于其他各组,但BCB+组、对照组以及控制对照组之间无显著差异.[结论]亮甲酚蓝染色筛选出的阳性卵母细胞具有较好的核成熟发育潜能,但并不能显著提高卵母细胞体外受精及孤雌激活后的胚胎发育率.