BCB染色液对绵羊不同直径卵泡卵母细胞的筛选作用

Screening of Sheep Oocytes with Different Follicle Diameters by BCB Staining

为探讨亮甲酚蓝（BCB）染色液对绵羊不同直径卵泡卵母细胞的筛选作用，对筛选后的卯母细胞进行孤雌激活，统计不同组间成熟率、卵裂率及囊胚率的差异。将卵泡直径分为〈2mm、2～5mm、〉5mm共3组，经26μmol／L的BCB染色90min后，结果显示：（1）直径〉5mm卵泡中的卵母细胞阳性率极显著高于5mm以下卵泡组（P〈0．01）；（2）卵泡直径〉5mm中BCB＋组卵母细胞孤雌激活后的体外培养效率显著高于其余2组；（3）不同试验组卵母细胞孤雌激活后获得的囊胚总细胞数在BCB＋组卵母细胞中，直径2～5mm组与〉5mm组间没有显著差异（P〉0．05），但极显著高于直径〈2mm组（P〈0．01）；BcB-卵母细胞在各组间没有显著差异（P〉0．05）；未经染色的卵母细胞中，直径2～5mm组与〉5mm组间没有显著差异（P〉0．05），但显著高于直径〈2mm组（P〈0．05）；此外，〈2mm的卵母细胞，BCB＋组极显著高于对照组和BCB一组（P〈0．01），在直径2～5mm组和〉5mm组，BCB＋组、对照组和BCB-组均差异极显著（P〈0．01）。以上结果说明，直径〉2mm以上卵泡的卵母细胞用BCB染色液筛选后更适合体外培养。

The aim of this paper was to study the effects of brilliant eresy efficiency of parthenogenetic activation （PA） with oocytes of different fol blue （BCB） staining on the cle diameters in sheep, and statistically analyze the maturation rates, cleavage rates and blastocyst rates in different groups. Base on the follicle diameter oocytes were divided into three groups：〈2 mm, 2--5 mm,）5 mm. Cumulus oocyte complexes （COCs） are stained with BCB （26 mol/L） for 90 min before in vitro maturation （IVM）. The results showed （1） positive rate of oocytes from follicles diameter larger than 5 mm group was significantly higher than that in Smm groups（P〈0.01） （2）after Oocytes were PA, the in vitro culture efficiency of oocytes from follicles diameter larger than 5 mm BCB＋ group were signif-icantly higher than others （P〈0.05） （3）the results of obtained total number of blastocyst cells from different experimental groups after PA showed that 2--5 mm group was not significant difference with5 mm group in BCB- group oocytes（P〈0.05）, but significantly higher than 〈2 mm group（P〈0.01） there was no significant difference in all BCB-- groups（P〈0.05）. In unstained oocytes, 2--5 mm group has no significant difference with 〉5 mm group （P〈0.05）, hut significantly higher than 〈2 mm group（P〈0.05）. Furthermore,in 〈2 mm group oocytes, BCB-＋ group was significantly higher than BCB--group and control group（P〈0.01） in 2--5 mm group and 〈5 mm group oocytes, there were significantly difference in all BCB＋ group, BCB-- group and control group（P〈0.01）. The results showed that oocytes from follicles diameter larger than 2 mm were appropriate for in vitro cultivation after BCB staining.