【摘要】 将从屠宰母牛卵巢采得的卵母细胞用含有LH或hCG的M199培养24～26小时,然后用经钙离子载体Ionophore A23187诱导获能的牛新鲜或冷冻精子进行授精处理。授精6～7小时后再用含有乳酸钠和EDTA的M199培养至48～60小时后观察受精及受精卵的发育情况。结果表明,卵子的受精率及受精卵的发育率与卵母细胞成熟用培养基、授精用精子(新鲜或冷冻)以及供卵牛的品种均有关系。将用含有hCG的M199培养成熟的卵子,以获能处理过的新鲜精子授精后,其受精率和受精卵子中单精子受精并发育为2～8细胞期胚的比率在蒙古牛为91.4％(265/200)和19.6％(52/265),在黑白花奶牛为69.7％(62/89)和4.8％(3/62)。更多还原

【Abstract】 The bovine ovaries were collected from local slaughterhouse and carried back to laboratory in warm saline within 8 hrs. Then the oocytes were aspirated from the small Graafian’s follicles ( 2~5 mm in diamiter ) . Only the morphologically normal oocytes with the compact cumulus were cultured in maturation medium M199 supplemented with 10% FCS and hCG or LH at the condition of 37℃ and 5 % CO2 in air. After 24~26 hrs, the matured oocytes with the expanded cumulus cells were inseminated with fresh or frozen-thawed semen capacitated with calcium ionophore A23187.Then the oocytes were transferred into the development medium M199 supplemented with FCS, sodium lactate and EDTA. After 48~60 hrs of insemination, the oocytes were fixed, stained and then examined with the phase-contrast microscope, The results indicated that the fertilization rate and the proportion of the oocytes developed to 2 ~ 8 cell stage were influenced by the maturation medium, semen and the breeds of the oocytes donor. In the case that the oocytes were matured in Ml99 + hCG and inseminated with fresh semen, the total IVF rate and the percentage of 2 ~ 8 cell stage embryos were 91.4% ( 265/290 ) , 19.6% < 52/265 ) in the oocytes collected from Mongolian yellow cow and 69.7% ( 62/89 ) ,4.8% ( 3/62 ) in Holtein cow, respectively.更多还原