卵巢运输温度及卵母细胞促成熟因子对山羊卵母细胞体外成熟和体外受精的影响

The Effect of Ovary Transfer Temperature and Maturation-promoting Factors on in vitro Maturation and in vitro Fertilization of Oocytes in Goats

探讨山羊卵母细胞体外成熟培养的影响因素,为优化山羊体外受精程序奠定基础。比较从屠宰场采摘的山羊卵巢运输温度和体外培养促成熟因子对卵母细胞体外成熟培养和体外受精效果的影响。结果表明,在运输温度接近室温(25±2)℃时的卵母细胞成熟率显著高于接近体温(36±2)℃时的运输温度(成熟率分别为53.8%,35.0%,P<0.01),两种运输温度的卵巢卵母细胞成熟培养后体外受精率无显著差异(分别为37.9%,36.6%,P>0.05)。与在体外成熟基础培养液(M1:M199 media+1μg/mL E2+10μg/mL FSH+10μg/mL LH+20%EGS)中进行成熟培养相比,在基础培养液中仅添加20 ng/mL EGF(M2)和另外添加10%GFF(M3)进行体外成熟培养的卵母细胞成熟率均有明显提高,其中成熟率在M1为53.8%,极显著低于M2的69.5%和M3的72.6%,P<0.01;M2和M3间差异不显著;卵母细胞成熟培养后体外受精率在三者间无明显差异分别为37.9%,39.5%和40.6%,P>0.05。山羊体外受精时从屠宰场采摘的卵巢宜在室温条件下进行运输,在体外成熟培养体系中加入EGF和GFF均可有效促进山羊卵母细胞的体外成熟,但EGF起关键作用。

The factors influencing maturation of goat oocytes in vitro were investigated to optimize the precedures of in vitro fertilization in goats.The experiments were carried out to compare the effect of deferent transferring temperature of ovaries collected from slaughtered goats and in vitro maturation-promoting factors on the efficiency of in vitro maturation and in vitro fertilization of oocytes.The maturation rate of oocytes come from the ovaries transferred at room temperature（25±2）℃ was significantly higher than that at body temperature（36 ± 2）℃（53.8%vs 35.0%,P〈0.01）.But there was no significant differences between the fertilization rates of mature oocytes come from the ovaries transferred at the two temperatures（37.9%vs 36.6%,P 〉0.05）.The maturation rate of oocytes cultured in basic media（M1：M199 media＋1 μg/mL E2 ＋10 μg/mL FSH＋10 μg/mL LH＋20%EGS）was significantly lower than that cultured in the basic media supplemented with 20 ng/mL EGF（M2）（69.5%vs 53.8%,P〈0.01）and in the basic media with 20 ng/mL EGF and 10%GFF（M3）（72.6%vs 53.8%,P〈0.01）.But the maturation rates of oocytes cultured in M2 and M3 were not significantly different（P〉0.05）.There were no significant differences among the fertilization rates of mature oocytes cultured in the three media（37.9%,39.5%and 40.6%,P〉 0.05）.The results indicated that the goat ovaries should be transferred at room temperature,both EGF and GFF can promote in vitro maturation of goat oocytes effectively,but EGF plays a key role.