金黄地鼠超排后未排卵泡卵母细胞的成熟度和受精能力的观察

Recovery and Utilization of Golden Hamster Ovary Oocytes After Superovulation

为了证实金黄地鼠超排后卵巢中未排卵卵泡卵母细胞的成熟程度和受精能力,用促性腺激素PMSG和HCG进行4次超排处理,其中选择12只超排效率低下的金黄地鼠,回收输卵管和卵巢卵母细胞,观察卵母细胞的成熟程度、体外培养成熟能力及体外受精能力。结果显示,输卵管中回收超排卵数平均为22.6±7.8枚/只,从超排后卵巢中回收卵丘扩展卵数平均为23.2±3.9枚/只,卵丘未扩展卵数平均为13.4±3.7枚/只。卵丘扩展卵经0.1%透明质酸酶消化,裸卵占86.1%,带放射冠卵占11.5±2.4%。裸卵中排出极体的成熟卵占卵丘扩展卵的80.4±2.3%,无极体裸卵占5.7±0.8%。经体外培养2h,无极体裸卵的成熟率达100%,带放射冠卵的成熟率仅为6.3±9.4%,而卵丘未扩展卵未见有成熟卵。人精子穿透试验显示,卵巢回收成熟卵的受精率44.1±6.0%,同超排卵的受精率45.4±4.7%比较,无显著差异(P<0.05)。从低超排率的金黄地鼠卵巢中可获取与超排卵数相近的成熟卵母细胞,且具有可利用于去透明带金黄地鼠卵子穿透实验(HOP)及相关发育生物学研究的潜能。

This thesis aims at characterizing the mature degree and fertilization ability of unovulation follecle in golden hamster after superovulation. The experiment workd on the basis of four times superovulation treatment with gonadotropin PMSG and HCG and obtained the oocytes in oviducts and unovulation follicle from 12 low superovulated hamster, so as to observe the mature degree and fertilization ability. The average amount of oocytes obtained from the oviducts was 22.6 7.8 per hamster, while the oocytes with expanded cumulus and the unexpanded ones from the unovulation follecle were respectively 23.2 3.9 and 13.4 3.7 per hamster. After the treatment with O. 1% hyaluronidase, 86.1% of the oocytes were naked and 11.5 2.4% had corona. Among the naked oocytes, 80.4 2.3% were matured with polar body while 5.7 0.8% had no polar body. After two hours cultures in vitro, the maturation rate reached 100% for naked oocytes with no polar body, 6.3 9.4% of the oocytes with corona and no maturation were observed from oocytes with a compact cumulus. The penetration test with human spermatozoa shows no significant difference （ P 〉 0.05 ） in the fertilization rate between matured oocytes （44.1 ± 6.0% ） and that of ovulated mature oocytes （45.4±4.7% ）. Similar number of mature oocytes could be retrieved from the ovaries as from that of oviducts, and those oocytes have the potentiality to be used for the zona - free hamster oocytes penetration test （HOP） and study for correlative development biology.