【作者】 周俊波；

【导师】 王杏龙；

【作者基本信息】 扬州大学 ， 动物遗传育种与繁殖， 2006， 硕士

【摘要】 本文主要采用两种不同组合的冷冻液(VSⅠ和VSⅡ)OPS法玻璃化冷冻不同期(GV、10h、IVM)的山羊卵母细胞,并对冷冻后的卵母细胞进行体外受精实验。主要结果如下:1.用两种不同组合的冷冻液VSⅠ(EG+DMSO)和VSⅡ(PROH+DMSO)冷冻各期(GV、10h、IVM)山羊卵母细胞。经用VSⅠ冷冻液冷冻后,GV期的形态正常率、成熟率、受精率分别为86.2%、25.3%、4.0%;培养10h后的分别为86.7%、25.6%、5.1%;IVM期的冻后形态正常率为87.8%、受精率为13.9%。经用VSⅡ冷冻液冷冻后GV期的形态正常率、成熟率、受精率分别86.9%、23.3%、2.7%;培养10h后的分别为88.0%、24.5%、3.3%;IVM期的冻后形态正常率为86.0%、受精率为13.5%。两组冷冻液对应各期的冻后形态正常率、成熟率和受精率差异均不显著(P>0.05)。2.无论是VSⅠ(EG+DMSO)还是VSⅡ(PROH+DMSO),山羊卵母细胞在GV期、体外培养10h后以及IVM期的冻后形态正常率上差异均不显著(P>0.05);GV期和体外培养10h后的卵母细胞的冻后成熟率差异不显著(P>0.05);但GV期和体外培养10h后卵母细胞的冻后受精率均低于IVM期卵母细胞,差异显著(P<0.05)。3.用获能精子和未获能精子,对成熟卵母细胞进行显微授精后,两组的形态正常率分别为81.5%和82.7%,差异不显著(P>0.05),获能精子组的受精率(28.3%)比未获能的精子组(18.8%)高,但差异不显著(P>0.05)。4.常规体外受精(IVF)的受精率(13.9%)比显微授精(ICSI)(28.3%)的低,两者差异显著(P<0.05)。说明ICSI技术相比较常规IVF更能够提高卵母细胞的受精率。更多还原

【Abstract】 In this paper, different stage of goat oocytes(GV、10h、IVM) were vitrified with two different concentration freezing media (VSⅠand VSⅡ) with the OPS method, after vitrified those oocytes were fertilized with IVF or ICSI. The main results were as follows:1. Vitrified different stages of goat oocytes(GV、10h、IVM) with two different concentration freezing media of VSⅠ(EG+DMSO)and VSⅡ(PROH+DMSO). After vitrified with VSⅠ, the morphologically normal rates、the IVM rates and the fertility rate of GV stage were 86.2%、25.3%、4.0%,respectively; and cultured 10h in vitro were 86.7%、25.6%、5.1%,respectively; the morphologically normal rate and the fertility rate of IVM stage were 86.0% and 13.5%, respectively. After vitrified, there was no significant difference in the morphologically normal rates、the IVM rates or the fertility rate between VSⅠ(EG+ DMSO) and VSⅡ( PROH+ DMSO) (P>0.05) .2. Either VSⅠ( EG+ DMSO) or VSⅡ( PROH+ DMSO), there was no significant difference in the morphologically normal rates、the IVM rates or the fertility rate of goat oocytes between each other of GV stage ,cultured 10h in vitro and IVM stage (P>0.05); there showed no significant difference between the IVM rates of GV stage and developed 10h in vitro; but the fertility rate of oocytes of GV stage and developed 10h in vitro were both lower than IVM, the difference was significance ( P<0.05).3. After microinsemination with capacitation and incapacitation sperms, the morphologically normal rates of maturate oocytes were 81.5%and 82.7%, respectively,更多还原