【作者】 许伍霞；

【导师】 薛立群；

【作者基本信息】 湖南农业大学 ， 动物遗传育种与繁殖， 2006， 硕士

【摘要】 本实验以屠宰厂采集的猪卵巢为材料，从中收集猪卵母细胞，进行玻璃化冷冻、体外培养及培养过程中NO含量变化以及不同大小卵泡NO含量测定研究，结果表明： 1．不同发育时期卵母细胞的冷冻效果有一定差异，未成熟的猪卵巢卵母细胞冷冻后形态正常率和存活率都低于正在成熟(培养24h)和已经成熟(培养44h)的卵母细胞，已经成熟的卵母细胞冷冻后损伤最小(形态正常率83.93％)，存活率最高(69.64％)。卵母细胞的NO含量随着成熟而逐渐降低。每个卵母细胞NO含量由0.480±0.026umol/L降到0.378±0.061umol/L。 2．卵泡φ＞6mm的卵母细胞冷冻后形态正常率，卵丘细胞扩展率，第一极体排出率比φ＜3mm的卵泡卵母细胞高。φ＞6mm卵泡卵母细胞形态完整率，卵丘细胞扩展率，第一极体排出率分别为84.78％、53.26％、35.70％，而φ＜3mm卵泡卵母细胞相应为60.19％、41.67％、9.26％，两者差异极显著(P＜0.01)。 3．不同载体对卵巢卵母细胞玻璃化冷冻效果不同，本实验表明麦管组不及GMP组，两者解冻后形态正常率分别为71.12％、50.04％；存活率分别为83.16％、62.50％，解冻后形态正常率差异极显著(P＜0.01)，存活率差异不显著(P＞0.05)。 4．用GMP法比较10％EG与10％DMSO预处理液，玻璃化冷冻液EFS30、EFS40对猪GV期卵母细胞冷冻效果，研究发现其解冻后形态正常率、培养24h卵丘细胞扩展率有不同，但其统计差异不显著(P＞0.05)。 5．在研究卵泡液对猪卵母细胞体外成熟的影响中发现，成熟液中添加10％PFF可以促进猪卵母细胞的体外成熟，使成熟率由60.06％提高到82.89％。 6．不同直径卵泡卵母细胞体外培养成熟44h后发现其卵丘细胞扩展率，第一极体排出率不同，φ为3—6mm最高，φ＞6mm次之，φ＜3mm最小，分别为73.26％、50％)；(62.79％、38.37％)；(54.48％、30.56％)，三者差异显著(P＜0.05)。大卵泡含NO最少，每个为2.618±0.341umol/l、中卵泡含NO次之，每个为3.840±0.321umol/L、小卵泡含NO最多，每个为4.597±0.685 umol/L、三者差异极显著(P＜0.01)。更多还原

【Abstract】 This experiment, with the discarded Porcine ovaries from the slaughterhouse as the experimental materials by carrying on the cryoperservation and raising Porcine Follicular Oocytes in vitro ,then measuring the content change of NO during the process and NO content of the follicles in different sizes ,indicates :1. The effect of freezing the oocytes in difffernt growth periods is different: the complete shape rates of unmatvire oocytes of porcine ovaries and the livability after frozen ,compared with the oncoming mature(raised for 24 hours )and already mature occytes (raised for 44 hours)are both lower ,while the mature oocytes will be damaged the least :the shape integrity rate is 83.90%;the livability is the highest (69.64%). the NO content of FoUicular Oocytes decreases gradually with increasing maturity of oocyte from 0.480±0.026 umol/L untile to 0.378±0.061umol/L for each FoUicular Oocytes.2. The rates of the shape integrity of oocytes of over 6mm in diameter and the oviferous tubercle cell expansion and First polocyte discharge, compared with those of occytes under 3mm in diameter are higher. when 0>6 mm, the mentioned -above rates of the FoUicular Oocytes are 84.78%、 53.26%、 35.70% respectively; when (?)<3 mm, it will be 60.19%、 41.67%、9.26% respectively . the difference is very significant(P<0.01).3. Different carriers for freezing ovary oocytes in glass will see different effects shown in this experiment, better in group strawpipe than the group GMP. After defrostation, the rates of shape integrity are 71.12% 50.04% ,while the livability are 83.16%、 62.50% respectively. The differences between are significant(P<0.01), the liabilities are not (P<0.05).4. Comparing the effects of the 10% EG and 10% DMSO pre-treatment liquids and the frozen glass liquids EFS30、 EFS40 on freezing oocytes in the period of Porcine GV with the method of GMP, we find after defrostation, the rates of shape更多还原