影响猪卵母细胞冷冻效果的若干因素  

Several factors affecting cryopreservation of porcine oocytes

作　　者：王昭凯[1] 吴华莉[1] 张德福[1] 刘东[1] 刘伟[2] 王少兵[2] 

机构地区：[1]上海市农业科学院畜牧兽医研究所 [2]南京农业大学动物科技学院,南京210095

出　　处：《上海农业学报》2008年第1期14-18,共5页Acta Agriculturae Shanghai

基　　金：上海市科技兴农重点攻关项目[沪农科攻字(2003)第14-1号];上海市科技兴农重点攻关项目[沪农科攻字(2005)第3-5号];上海市农业科学院青年基金(2003-09)资助

摘　　要：以屠宰场废弃卵巢卵母细胞为研究对象,探讨了不同冷冻保护剂、冷冻载体、成熟时期、去脂处理和不同受精方法对冻后卵母细胞存活和发育的影响。结果表明:渗透性保护剂的毒性大小依次为EG<GL<DMSO;一步法冷冻MII期卵母细胞,以EFS40的效果最好,显著优于其他玻璃化液组;在玻璃化液与卵母细胞的作用时间上,30 s和1 min组之间无显著差异,但显著好于2 min处理时间组;以GLT(55.8%)为冷冻承载工具,效果要好于OPS(29.5%)和GMP(13.4%)法,且差异显著;随着卵母细胞的成熟发育进程,卵母细胞的抗冻能力呈上升趋势;与对照组(53.6%)相比,CB(55.6%)和CB离心处理(54.5%)能部分提高卵母细胞的冻后存活力,去脂处理并不能提高卵母细胞的存活率;冷冻MII期卵母细胞经体外受精途径未能获得卵裂,经ICSI途径,获得6.1%的冻后卵母细胞发育率。Porcine ovaries from a slaughterhouse were used as a test material, and the effects of different cryoprotectants, cryopreservation tools, in vitro maturation stages, treatments of CB＆ Lipid removal and fertilization methods on the survival and development of vitrified porcine oocytes were investigated. The results showed that the ascending order of the toxicities of the osmotic cryoprotectants was EG〈GL〈DMSO. In the one-step vitrification of the MII-stage oocytes, the vitrification solution of EFS40 got the best result, and was significantly better than other vitrification solutions. For the time of treating the ooctyes with vitrification solutions, both 30 s and 1 min groups were not significantly different but significantly better than the 2 min group. Using GLT （55.8%） as cryopreservation tool got a significantly higher survival rate than using OPS （29.5%） and GMP （13.4%） methods. With the development of oocytes in the process of in vitro maturation, the anti-freezing ability of ooctyes increased. Compared with the control （53.6%）, CB （55.6%） and Centrifugation ＋ CB （54.5%） treatments could partly increase the survival rate of MII-stage oocytes after freezing, but lipid removal did not improve the survival rate of oocytes. Any cleavage from the vitrified MII-stage oocytes could not be got by in vitro fertilization method, but 6.1% cleavage rate could be got by the way of ICSI.

关 键 词：冷冻保存 玻璃化 猪 体外受精 显微受精 

分 类 号：S858.28[农业科学—临床兽医学] S814.6[农业科学—兽医学]