人腔前卵泡分离及培养  

Isolation and culture of human ovarian preantral follicles

作　　者：施旭东[1] 孙贝加[2] 谭笑梅[3] 王公金[4] 陈华[3] 孔庆■ 陈杰[1] 范必勤[4] 

机构地区：[1]南京农业大学动物医学院动物生物生理化实验室,210095 [2]东南大学医学院徐州附院,221009 [3]南京市妇幼保健院,210004 [4]江苏省农科院畜牧兽医研究所胚胎工程实验室,江苏南京210014 [5]徐州医学院病理教研室,江苏徐州221000

出　　处：《生物技术》2001年第4期13-17,共5页Biotechnology

摘　　要：采用机械吹打、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。将分离得到的腔前卵泡在含FSH0 5IU/mL、1 0IU/mL和 2 0IU/mL的培养液中培养 ,检测其E2 分泌量。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)。得到的腔前卵泡在FSH为 0 5IU/mL、1 0IU/mL和 2 0 1IU/mL的培养液中培养 ,所分泌的E2 分别为 10 86pg± 4 11pg、31 5 5pg± 9 34pg和4 3 82pg± 18 76pg ,较之对照组的 4 99pg± 2 0 9pg有显著性差异 (P <0 0 1) ,E2 的分泌量与FSH浓度呈剂量依赖性关系。FSH有促进腔前卵泡分泌E2To establish a method for isolation and collection of human ovarian preantral follicles,four isolation methods,mechanical pipetting, collagenase digestion, microdissection and the combination of collagenase digestion and microdissection were performed and compared. The isolated preantral follicles were cultured in media containing FSH with the concentration of 0.5 IU/mL,1.0 IU/mL and 2.0 IU/mL.The amount of E 2 secreted by preantral follicles was measured. In comparison with mechanical pipetting and collagenase digestion, more follicles (P<0 01) and various follicles:primordial,primary and secondary follicles were obtained by using the method of collagenase digestion combined with microdissection despite its time consuming(P<0.01). The amount of E 2 secreted by preantral follicles cultured in vitro supplemented FSH was 10.86±4.11pg,31.55±9.34pg and 43.82±17.86pg per follicle for the concentration of 0.5 IU/mL, 1.0 IU/mL and 2.0 IU/mL respectively. Compared with the amount of E 2 secreted by control group, 4.99±2.09 pg,there were significant differences (P<0.01) between groups with FSH and control group and a dose-dependent relation. Collagenase digestion combined with microdissection is an efficient method for isolation and collection of follicles from human ovaries. FSH increases the amount of E 2 secreted by preantral follicles.

关 键 词：人 腔前卵泡 显微解剖 分离 体外培养 FSH E2 

分 类 号：R321－33[医药卫生—人体解剖和组织胚胎学]