人类卵巢卵泡显微解剖分离方法研究

Study on the microdissection method for isolation of human ovarian follicles

目的 建立人卵巢卵泡分离提取的方法。方法 尝试机械吹打、胶原酶消化、镜下显微解剖及胶原酶消化加镜下显微解剖 4种卵泡提取方式并相互对比。观察不同年龄阶段卵巢的卵泡数量。结果 卵巢中的卵泡数随年龄的增加而减少。与机械吹打、胶原酶消化相比 ,胶原酶消化加镜下显微解剖法不仅提取卵泡多 (P <0 0 1) ,而且可以得到原始、初级、次级各级卵泡 ,但操作时间较长 (P <0 0 1)。随机抽取各组卵泡体外培养 ,48h后观察其结构正常 ,并能产生E2 。结论 胶原酶消化加镜下显微解剖是人卵泡分离提取的有效方法。

Objective To establish a method for the isolation and collection of human ovarian follicles.Methods Four isolation methods,including mechanical pipetting,collagenase digestion,microdissection and the combination of collagenase digestion and microdissection,were performed and compared.The quantity of follicles from various phases of age was observed.Results The quantity of follicles decreased with the increasing age.In comparison with mechanical pipetting and collagenase digestion,more follicles(P<0 01) and various follicles,including primordial,primary and secondary follicles,were obtained by using the method of collagenase digestion combined with microdissection despite its time consuming(P<0 01).Follicles in each group were selected randomly and cultured in vitro.After 48h,follicles maintained normal morphology under inverted microscope and produced detectable E 2.Conclusion Collagenase digestion combined with microdissection is an efficient method for isolation and collection of follicles from human ovaries.