不同冷冻载体对牛未成熟卵母细胞玻璃化冷冻效果的影响  

Effect of different cryopreservative carriers on vitrification of bovine immature oocytes

作　　者：裴旭哲 马德全 张凡[1,2] 武花 郭先飞 李晓霞[1,2] 栗颖华[1,2] 禹学礼[1,2] 

机构地区：[1]河南科技大学动物科技学院,河南洛阳471023 [2]河南省草食动物工程重点实验室培育基地,河南洛阳471023 [3]洛阳市畜牧良种繁殖推广中心,河南洛阳471013

出　　处：《畜牧与兽医》2017年第5期58-62,共5页Animal Husbandry & Veterinary Medicine

基　　金：国家自然科学基金(31172207和31201801);河南省教育厅科学技术研究重点项目(14A230015)

摘　　要：为了研究不同冷冻载体制备方法及其对牛未成熟卵母细胞发育能力的影响,以牛未成熟卵母细胞为实验材料,分别探究开放式拉长细管(OPS,open pulled straw)和玻璃微管(GMP,glass micropipette)的制备方法,并以OPS、GMP和冷冻叶片为冷冻载体玻璃化冷冻牛未成熟卵母细胞,解冻后经体外成熟培养和体外受精,统计形态正常率、成熟率、卵裂率及囊胚率。结果显示:以简易小酒精灯为热源,以细管为原材料可以制备出适用的OPS冷冻载体;以酒精喷灯为热源,以细玻璃管为原材料可以制备出适用的GMP冷冻载体。OPS和GMP组形态正常率分别为(74.3%±1.8)%和(72.5%±2.6)%;无显著差异(P>0.05),上述二组均显著低于冷冻叶片组(82.1%±1.3)%,P<0.05,但三组间成熟率、卵裂率和囊胚率均无显著差异(P>0.05)。研究表明,分别以简易小酒精灯和酒精喷灯为热源,以细管和细玻璃管为原材料可以成功制备出OPS和GMP载体;以OPS、GMP和冷冻叶片为冷冻载体均可成功地玻璃化冷冻牛未成熟卵母细胞。The aim of this study was to explore the fabrication methods of different cryopreservative carriers and their effect on vitrification of bovine immature oocytes. Firstly, open pulled straw （OPS） and glass micropipette （GMP） were made successfully by hand. Then, bo- vine cmnulus oocyte complexes （COCs） were vitrified using OPS, GMP and Cryoleaf as carriers respectively. After warming, the oocytes were maturated, fertilized and cultured in vitro. The rates for normal morphology, maturation, cleavage, and blastocyst were determined. Re- sults showed that OPS could be made of straws using simple small alcohol lamp. GMP could be made of thin glass tube using alcohol burner. The normal morphology of cryoleaf group （ 82. 1 ± 1.3 ） % was significantly higher than OPS （ 74. 3 ± 1.8 ） % and GMP （ 72. 5 ± 2. 6） % group （P〈0. 05 ） , however, there was no significant difference between OPS and GMP group （P〉0. 05）. Furthermore, there were no significantdifferences in the maturation rate, cleavage rate and blastocyst rate among three groups （P〉0. 05）. Together, bovine immature oocytes can be vitrified effectively by OPS, GMP and Cryoleaf.

关 键 词：牛 未成熟卵母细胞 冷冻载体 玻璃化冷冻 

分 类 号：S823.3[农业科学—畜牧学]