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

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.