牦牛卵母细胞的体外成熟

In vitro maturation of yak oocytes

以屠宰场牦牛卵巢为材料,比较了抽吸加切割法和抽吸法2种卵母细胞离体采集方法的效率和5种成熟培养液的培养效果,并研究了卵泡位置、形态对卵母细胞体外成熟的影响。结果表明:在牦牛乏情期,平均每个卵巢用抽吸加切割法回收卵数极显著高于抽吸法(9.33±4.30VS4.70±2.62,P<0.01),可用卵数也极显著高于抽吸法(5.63±4.19VS4.37±2.32,P<0.01)。将牦牛卵丘卵母细胞复合体(COCs)分别置于5种成熟培养液中培养,其中M199(缓冲体系为Earles盐)+10% FBS+5.0mg/LLH+1.0mg/LE2+双抗(青霉素100IU/mL和链霉素100mg/L)的成熟液效果最好,成熟率为81.33%,卵裂率为49.33%。来自卵巢表面卵泡的COCs的成熟率和卵裂率均高于来自卵巢内卵泡的COCs(分别为81.33%VS69.33%,P>0.05;49.33%VS34.67%,P<0.05)。来自明亮卵泡的COCs的成熟率和卵裂率均极显著高于来自浑浊卵泡的COCs(分别为81.33%VS33.33%,P<0.01;49.33%VS3.33%,P<0.01)。

This experiment was carried out to compare the efficiency of two kinds of oocytes collection method and the effects of five types of maturation media, to study the effects of what place in ovary and morphological character of follicles on oocytes maturation in vitro. The results were as follows ：during yak＇s anestrus ,both No. of oocytes and No. of useful oocytes from each ovary by scoring the ovarian surface with a set of surgical blades followed aspiration of follicular oocytes were significantly higher than those by only aspiration of follicular oocytes （ 9.33 ± 4.30 VS 4.70± 2.62,5.63± 4.19 VS 4.37± 2.32,P〈0. 01 ）. Yak＇s cumulus-oocyte complexes （COCs） were cultured in five maturation media respectively,with the maturation rate of 75.56%, 71.11%,81.33%, 77.33% ,77.78%, respectively and the cleavage rate of 42. 22% ,33.33% ,49.33% ,46.67% ,42.22%, respectively. The maturation medium of M199（with Earle＇s salts）＋10% FBS＋ 5.0 mg/L LH＋1.0 mg/L E2＋antibiotics （100 IU/mL penicillin and 100 mg/L streptomycin） is the best. Cleavage rates of COCs from surface follicles were significantly higher than that from interior follicles （49. 33 % VS 34.67 %, P〈 0.05）, but maturation rates didn＇t exist significant difference between them （81.33% VS 69.33% ,P〉 0. 05）. Both maturation and cleavage rates of the COCs from the pellucid follicles were significantly higher than those from the turbid follicles（81.33% VS 33.33%,49.33% VS 3.33%,P〈0.01）.