【摘要】 从牛屠体卵巢采得卵母细胞后,置于CO₂培养箱中作24小时成熟培养。在成熟培养液中分别添加0％、10％、20％、30％牛新鲜卵泡液（BFF）或灭活卵泡液（hBFF）。用分层Percoll液将解冻精子离心洗涤后,悬浮于含有10μg/ml肝素的修正Tyrode液滴中,将培养成熟的卵子移入后作授精培养。20小时后,固定、染色一部分卵子,镜检其受精情况,其余卵子移入发生培养基与颗粒膜细胞作混合培养。试验结果表明,新鲜卵泡液添加组与对照组比较,在受精率、多精子侵入率以及受精后的早期卵裂率方面均无显著差异。在囊胚发生率方面,30％添加组（29.8％）显著高于对照组（13.5％）（P<0.05）;极显著地高于10％添加组（11.5％）（P<0.01）。但是,在成熟培养基内添加灭活卵泡液后,各组间均无显著差异。因此,对牛卵母细胞作成熟培养时,添加适量新鲜卵泡液,可以提高受精后的发育潜能,提高囊胚的发生率。更多还原

【Abstract】 The follicular oocytes collected from slaughterhouse ovaries were matured for 24 h. at 39℃ under 5% C02 in air with high humidity under paraffin oil. The maturation media were supplemented with 0%, 10%, 20%, 30% bovine follicular fluid (BFF) or heat-inactivated bovine follicular fluid (hBFF), respectively.Frozen-thawed semen was layered on bilayered percoll solution (30% and 45% of 2 ml each) and centrifuged. The pelleted spermatozoa were resuspended with defined medium including 10μg/ml heparin and were co-cultured with matured oocytes at 39℃ under 5% C02 in air. Some of the eggs were fixed and stained at 20h. after insemination, and examined by phase microscopy. The others were transferred into droplets of developmental medium for co-culture with granulosa cells.The addition of BFF to the maturation medium showed no significant difference in the rates of fertilization, polyspermic eggs and early cleavaged eggs as compared with that of the control. The rate of blastocysts in 30% BFF group (29.8%; 28/94) were significantly higher than that of 10% BFF group (11.5%; 10/87) (P<0.01) and control (13.5%; 12/89) (P<0.05), but no significant difference were found as compared with that of 20% BFF group (24.7%; 23/93). The addition of hBFF to the maturation medium showed no significant difference as compared with the control.更多还原