不同分离方法及培养方式对绵羊腔前卵泡发育能力的影响  

Effect of Different Isolation Techniques and Culture Methods on Developmental Competence of Sheep Ovarian Preantral Follicles

作　　者：常迪[1] 李贺娟[1] 尹艳云[1] 陈利平[1] 刘云海[1] 郭勇[1] 倪和民[1] 

机构地区：[1]北京农学院动物科技学院,北京102206

出　　处：《中国农学通报》2010年第18期27-31,共5页Chinese Agricultural Science Bulletin

基　　金："十一五"863计划"家畜卵泡高效诱导发育技术研究与应用"(200AA101003);国家自然基金面上项目"绵羊体外胚胎妊娠时MUC1;GLYCAM1和ISG15对子宫容受态影响的研究"(30972228)

摘　　要：研究目的在于分析用不同的方法分离绵羊腔前卵泡以及用不同的培养方式对分离得到的腔前卵泡进行培养,对于绵羊腔前卵泡存活和发育能力的影响。试验分2部分进行:试验1为分离卵巢皮质,分别用显微分离法和胶原酶法分离腔前卵泡。试验2是在试验1的基础上,将较优方法获得的腔前卵泡分别进行单独培养和群体培养。结果显示,显微分离法与胶原酶法相比,在获得的腔前卵泡数量上差异不显著,但显微分离法获得的腔前卵泡基膜完整率显著高于胶原酶法(P<0.05)。获得的腔前卵泡使用单独和群体两种方式进行培养,其直径增加值在第3天和第6天差异不显著,但单独培养法在第3天和第6天的存活率显著高于群体培养法(P<0.05),且有卵泡腔形成。研究显示使用显微分离法分离腔前卵泡与胶原酶法相比具有优势,相对简单、廉价,并且对卵泡的损害很小。经显微分离法获得的腔前卵泡,使用单独培养法进行培养能够促进卵泡的生长发育。The present study was aimed to analyze different techniques for isolating the sheep preantral follicles （Experiment I） and their different subsequent in-vitro culture methods of the isolated preantral follicles （PFs, Experiment II）. In Experiment I, the ovine ovarian cortical pieces were separated, and PFs were isolated by different techniques. In Experiment II, the isolated PFs from the superior technique based on the outcome of Experiment I, were divided into two groups for culture in vitro in a single follicle independently or in groups. The results showed that there were no significant differences between the micro-dissected way and collagenase-treated one on the isolated PFs number, but the survival rate of micro-dissected PFs was significantly higher than that of the group treated by collagenase （P 0.05）. In addition, in Experiment II, there were no significant differences between those two methods on increase of follicular diameters, but the survival rate of PFs cultured in vitro in single follicle independently, was significantly higher than that of PFs cultured in groups（P 0.05）, as well as their formed follicular antrum. Collectively, it can be concluded that the micro-dissected technique is superior to collagenase one, and PFs cultured in vitro independently can improve the developmental competence of those preantral follicles.

关 键 词：绵羊 腔前卵泡 分离 培养 发育能力 

分 类 号：S814.8[农业科学—畜牧学]