猪胚胎的OPS玻璃化冷冻  

Piglets born after OPS vitrification of zona pellucida-intact embryos

作　　者：张德福[1] 刘东[1] 吴华丽[1] 郑筱峰[2] 王昭凯[3] 王少兵[3] 

机构地区：[1]上海农业科学院畜牧兽医研究所上海市农业遗传育种重点实验室,上海201106 [2]南京农业大学动物医学院,江苏南京210095 [3]南京农业大学动物科技学院,江苏南京210095

出　　处：《中国兽医学报》2007年第6期947-949,共3页Chinese Journal of Veterinary Science

基　　金：上海市科技兴农重点攻关资助项目(沪农科攻字(2003)第14-1号)

摘　　要：以广西巴马小型猪为供体，采用超数排卵技术，采集5～6日龄具有完整透明带的胚胎（囊胚／桑葚胚），采用二步法OPS（Open pulled straw）玻璃化冷冻技术进行保存，即胚胎首先在冷冻液1（TCM199＋20％FBS＋10％EG＋10％DMSO）中平衡3min，然后立即转入冷冻液2（TCM199＋20％FBS＋20％EG＋20％DMSO＋0．4mol／LSUC）中并在1min内装管（每管含2～6枚胚胎），直接投入液氮保存；3个月后解冻移植给8头受体母猪（每头移入25～26枚胚胎），其中1头怀孕产仔（8头活仔），获得猪胚胎超低温（-196C）冷冻后代。In Bama-mini-pigs, five to six days post-insemination,morula and blastcysts were collected together after uterine. The purpose of this study was to vitrify zona pellucida-intact morula or blastcysts with OPS （open pulled straw） technology and obtain piglets after transfer in China. Blastocyst/morula were vitrified after a twostep equilibration in ethylene glycol,dimethyl sulfoxide and sucrose in Hepes-buffered TCM199＋20%FBS. 2-6 blastocyst/morula were loaded into OPS and plunged into liquid nitrogen. At embryo warming ,a three-step dilution with decreasing concentrations of sucrose was applied. In each of 8 recipients,25-26 blastocyst/morula were transferes surgically. The pregnant recipient farrowed 8 alive piglets. Although survival rates are still compromised ,OPS technology is therefore appropriate to cryopreserve porcine morula or blastcysts with intact zona pellucida.

关 键 词：猪 冷冻胚胎 玻璃化冷冻 OPS法 

分 类 号：S814.8[农业科学—畜牧学]