玻璃化快速冷冻仪在猪MⅡ期卵母细胞玻璃化冷冻中的应用  

Application of Vit-Master to vitrification of porcine MII oocytes

作　　者：徐利[1,2,3] 戴建军[1,2] 张廷宇[1,2] 吴彩凤[1,2] 李维杰[4] 张树山[1,2] 顾晓龙[1,2] 李翔[1,2] 韩晓[1,2,3] 李芳芳[1,2,3] 

机构地区：[1]上海市农业遗传育种重点实验室动物遗传工程研究室,上海201106 [2]上海市农业科学院畜牧兽医研究所,上海201106 [3]上海海洋大学水产与生命学院,上海201306 [4]上海理工大学生物热科学研究所,上海200093

出　　处：《上海农业学报》2014年第1期17-20,共4页Acta Agriculturae Shanghai

基　　金：上海市科委农业科技成果转化项目(123919N0700;1333919N1700);国家自然科学基金(31372315);上海市农业科学院青年科技发展基金(2011-07);国家转基因生物新品种培育科技重大专项(2008ZX08006-005;2009ZX08006-014B)资助

摘　　要：为了提高猪成熟卵母细胞玻璃化冷冻效果,应用玻璃化快速冷冻仪(Vit-Master)进行猪成熟卵母细胞的冷冻,以观察其冷冻效果。结果显示:浆状液氮降温速率[(21 345±l768)℃/min]远远高于普通液氮降温速率[(11 982±1 936)℃/min];各组冻后形态完整率无显著差异;冻后存活率为:使用玻璃化快速冷冻仪的不同浓度保护剂组[A':(57.87±10.66)%、B':(76.5±10.33%)、C':(79.07±14.90)%和D':(81.94±6.98%)]均分别高于对应的不使用玻璃化快速冷冻仪组[A:(49.73±2.91)%、B:(60.63±12.45)%、C:(71.33±14.38)%和D:(78.50±7.40)%],但均无显著差异,B'和C'组均高于C和D组,但差异不显著,A和A'组均显著低于B'、C'和D'组。因此,通过使用Vit-Master冷冻装置提高冷冻速率,可以降低冷冻保护剂使用浓度并提高冻后存活率。In order to improve the vitrification efficiency of matured porcine oocytes, the Vit- Master vitrification machine was used to freeze the in vitro matured porcine oocytes. The results showed that the cooling rate of slush liquid nitrogenE（21 345 ± I 768） °C/mini was much higher than that of normal liquid nitrogen[-（11 982- 1 936）°C/mini. All the groups had no significant differences in post-thaw morphological integrity. The post-thaw survival rates of the groups by using the Vit- MasterEA＇：（57.87 ± 1（1.66）%,B＇： （76.5 ± 10.33）%, C＇： （79.07 ± 14.90）% and D＇： （81.94 ± 6.98）%] were higher than those of the corresponding groups without the Vit-MasterEA： （49.73 ± 2.91）%,B：（60.63± 12.45）%,C：（71.33 ± 14.38）% and D：（78.50 ±7.40）%],but there were no significant difference among them. The survival rates of Groups B＇ and C＇ were higher than those of Groups C and D,but the differences were not significant. The survival rates of Groups A and A＇ were significantly lower than those of Groups B＇ ,C＇ and D＇. From the above results it was concluded that the Vit-Master application could increase the cooling rate, reduce the cryoprotectant concentrationand improve the post^thaw survival rate of oocytes.

关 键 词：猪 卵母细胞 玻璃化冷冻 玻璃化快速冷冻仪 降温速率 半麦管 

分 类 号：S814.8[农业科学—畜牧学]