摘要
从屠宰场收集水牛卵巢223个,由2—8毫米直径的卵泡中回收卵母细胞302枚,择其卵丘细胞紧密、细胞质均匀者265枚用于试验,于不同条件下体外培养24—27小时,成熟率为16.7—80％。水牛冷冻精液经解冻作精子上游和获能处理,对卵母细胞授精后培养6小时,休外受精率10—60％,移入培养液中继续培养获得一枚4-细胞期胚胎。另将受精卵移入家兔输卵管中作体内培养60小时,回收23枚受精卵中有3枚发育为16—32-细胞胚。此即表明,水牛卵母细胞可体外培养成熟和受精,并在体外或异种动物体内进行卵裂。 卵母细胞成熟培养结果,在A成熟培养液中,5％CO_2空气比空气环境培养的成熟率高,分别为58.3％和16.7％;在B成熟培养液中,37.5℃的培养效果优于39℃,成热率分别为80％和65.7％;在不同培养液中培养的卵细胞成熟率无显著差异;直径大于5毫米的卵泡卵母细胞,成熟率高于直径小于5毫米卵泡的卵母细胞,分别为79.3％和58.5％,受精率则相反,分别为23.5％和60％。
Three hundred and two oocytes were obtained from follicles (2-8mm in diameter) of 223 water buffalo ovaries collected at a slaughterhouse. T wo hundred and sixty-five oocytes with compact cumulus cells were selected to mature in vitro under specific conditions for 24 to 27 hrs, and resulted in a maturation rate of 16.7-80%. The fertilization rate was 10-60% 6 hrs after insemination with thawed frozen semen capacitated in vitro. One of the six fertilized ova normally developed to 4-cell stage 48 hrs after culture] in vitro. After 6 hrs exposed to sperm, 38 oocytes were transferred to the ligated oviducts of rabbit injected with HCG. After cultivation for 60 hrs in vivo, three of twenty-three oocytes recovered developed into 16- to 32-cell stages. The results indicated that follicular oocytes of water buffalo could be matured and fertilized in vitro, and normal cleavage occurred when they were cultured in vivo and in vitro.
The results also showed that maturation rates of oocytes cultured in air containing 5% CO2 were higher than in ambient air (58.3% vs 16.7%) and were higher for those cultured at 37.5癈 than at 39癱(80% vs 65.7%). There was no significant difference in the maturation rate of oocytes cultured in different media. The oocytes collected from follicles^ 5 mm in diameter had higher maturation rates than those from follicles <5 mm ( 79.3% vs 58.5% ) , but fertilization rates were higher for those from follicles <5 mm than those from follicles>5 mm (60% vs 23.5%).
出处
《江苏农业学报》
CSCD
1990年第3期25-30,共6页
Jiangsu Journal of Agricultural Sciences
基金
国家863高技术课题
关键词
水牛
体外受精
卵母细胞
体对成熟
Water buffalo
Follicular oocyte
in vitro maturation
in vitro fertilization
