恒河猴(Macaca mulatta)卵细胞采集及体外受精的研究  

A Study on Oocyles Recovery and Fertilization of Rhesus Monkey ( M.mulatto ) in Vitro

作　　者：陈乾生[1] 欧阳子焯[1] 黄丽碧[1] 刘晓明[1] 卢光琇[2] 陆长福[2] 李波[2] 刘展环[3] 

机构地区：[1]华南濒危动物研究所 [2]湖南医科大学生殖工程中心 [3]中国科学院动物研究所

出　　处：《中国实验动物学报》1993年第1期26-31,共6页Acta Laboratorium Animalis Scientia Sinica

摘　　要：对13头成年母猴,用黄体酮+前列腺索(PGF_(2a))处理,诱发月经周期,并用三种不同的处理方案,注射外激性促性腺激素诱发等超数排卵。对171个大滤泡(直径≥4mm)实行外科手术采卵,滤泡穿刺吸卵共获147个卵细胞,平均每头猴采卵11.31±6.47个,卵细胞回收率为85.96％。精液用直肠电刺激法采集,精子经TALP培养基洗涤二次,并在含10％灭活猴血清的TALP液中,加入1mM CAMP和lmM咖啡因,预孵育6～3h。卵细胞在含10％灭活猴血清的TALP或Ham'sF-10培养基中作体外成熟的培养,经约4～12h孵育,卵细胞体外成热率为28.57％。成熟的卵细胞(n=42,中期Ⅱ)和已预孵育的精子一起12～24h,并有部分在前次输精后5～12h再次输精,在42～56h后有9个受精卵继续发育至2细胞期和桑椹期阶段,体外受精率为35.71％(15/42)。The men trual cycl s were induced with prog steron and pr stagl ndin F20 in thirteen adult f male monkeys (5~10 years old, body weight 7~9kg) . They has received the treatment of exog nous gonadotr phin to indue superovulation in three experimen talp rocedures, 147 oocytes were coll ct d by penetrated aspiration, the average of oocyte collection is 11.31± 6.47, 171 large follicles ( diameter≥4.0mm ) were talen by surgical operation. The recovery rate of oocyte i s 85.93%. Semens were collected by electro-ejaculation, ths spermatozoa were washed with TALP and pre-incubated in TALP medium containing 10% heat inactivated l omologous serum, lmM cAMP and 1mM caffeine for 6~3h . The oocy tes were cultivated in TALP or Ham's F^10 medium containing 10% heat inactivated homologous serum for maturation in vitro, after 4~12 h of incubation the maturity rate is 28.57%. The mature oocytes(M Ⅱ , n = 42)were co-cultivated with capacitated spermstrozoa for 12~24 h, and part of these were again inseminated 5~12 h late.As result, fifteen of these oocytes were fertilized and 9 of these were developed to 2~cell or morula embryos for 42~56 h of cultivation.The fertilization rate was 35.71% ( 15/42 ), The result suggests that oocytes of Rhesus monkey the cytoplasmic maturation in vitro were required for longer time. The fertilzation rate of oocytes would be increased by the method of double insemination.

关 键 词：卵细胞 诱发 体外受精 血清 精子 恒河猴 孵育 采卵 体外成熟 培养基 

分 类 号：Q959[生物学—动物学] R321[医药卫生—人体解剖和组织胚胎学]