水牛小腔卵泡分离方法的初步研究

Studies on Isolation of Buffalo Small Antral Follicles

本研究比较了不同的分离方法和酶作用的不同时间,以及不同的卵巢表面状态等因素对水牛小腔卵泡分离效果的影响,以期建立有效的水牛小腔卵泡分离体系。结果发现,采用机械与酶结合的方法分离水牛小腔卵泡时,平均每个卵巢获得5.64个小腔卵泡,显著高于机械法分离平均每个卵巢获得的3.12个小腔卵泡数(P<0.05);当胶原蛋白酶作用12或15min,每个卵巢平均获得的小腔卵泡数均显著高于5和10min处理组(P<0.05),两组之间没有显著差异(P>0.05),但当消化时间延长到20min时,卵泡膜被酶消化而破裂,没法分离到小腔卵泡;从表面小于2mm卵泡且无黄体的卵巢分离获得的小腔卵泡平均数显著高于表面无可见卵泡且有黄体的卵巢组的平均数(7.50和2.25,P<0.05)。以上结果表明,选用表面无小于2mm卵泡且无黄体的卵巢,采用机械与酶结合的方法,酶作用时间为12～15min分离水牛的小腔卵泡,可获得的小腔卵泡数量最多。

This article was conducted to study the factors of isolation buffalo small antral follicles,so that to establish the effective isolation method of buffalo small antral follicles.The results showed that the method of mechanical combine with enzyme harvested significantly more small antral follicles（5.64） than mechanical method（3.12,P〈0.05）.The average number of small antral follicles harvested from ovary was increased as enhancing the digesting time of enzyme,and the groups of 12 min and 15 min were significantly higher than that of digesting for 5 and 10 min（P〈0.05）,but when digesting time of enzyme was prolonged to 20 min,the theca of follicles was broken out that could not isolate the intact small antral follicles.Most small antral follicles harvested from ovary which surface had less than 2 mm follicles but no corpora luteum significantly higher than that of ovary which surface had corpora luteum but no follicles（7.50 and 2.25,P〈0.05）.These results indicated that selecting the ovary which surface had less than 2 mm follicles but no corpora luteum and adopting the method of mechanical combine with enzyme to digest 12 to 15 min was fit for isolating the buffalo small antral follicles.