山羊胚胎简易冷冻试验

SIMPLIFIED QUICK FREEZING OF GOAT EMBRYOS

用RPE冷冻器冷冻20枚山羊桑椹胚和囊胚。冷冻液为1M甘油杜氏磷酸盐缓冲液(PBS)。胚胎装于0.25ml塑料细管内,0℃时置入冷冻器。以1℃/分降温,-6.5～-7℃诱发结晶,再以0.5℃/分(0.3～0.8℃/分)降温至-30℃,然后投入液氮保存5～12天。35℃水浴解冻。20枚胚胎冻后形态正常率为65％(13/20)。将13枝形态基本正常的胚胎移植给5只受体羊,结果4只受体羊妊娠,产羔7只,冻胚存活率为35％(7/20)。除1只冻胚羔产下后90分钟死亡外,另6只冻胚羔生长发育正常。

Twenty goat morulae and blastocysts were frozen in 1 M glycerol/ Dulbecco's phosphate-buffered saline supplemented with 10% heat-tre- ated fetal calf serum in the RPE freezer. Embryos were loaded into 0.25 ml plastic straws that were placed into the freezing apparatus at 0 ℃. The cooling rate was about 1 ℃/min from 0 ℃ to -5.5℃. They were seeded between -6.5℃ and - 7 ℃ and cooled from -7 ℃ to -30℃ at the rate of 0.5℃/min ( 0.3-0.8℃/min ) ,then plunged into liquid nitrogen and stored for 5 -12 days. Thawing was in a water bath at 35 ℃. After thawing, the percentage of normal embryos was 65%(13/20). Thirteen normal embryos were transferred into the uteri of five recipients, four of which became pregnant and produced seven kids. The embryo survival rate was 35% (7/20) . Except for one frozen-embryo kid that died 90 minutes after birth, kids were normal in growth and development.