小鼠胚胎直接投入液氮冷冻保存试验

室温下,将292枚小鼠胚胎在含128.9g/L甘油PBSS(含20％犊牛血清的磷酸盐缓冲液)和含193.2g/L甘油+85.6g/L蔗糖PBSS内分别孵育片刻,装管后直接投入液氮冷冻保存。冻胚快速解冻后,用含342.3g/L蔗糖PBSS脱去甘油,用PBSS清洗后镜下观察评定,胚胎回收率为86.9％(254/292),胚胎形态正常率为77.1％(196/254)。其中部分桑椹胚经体外培养,囊胚发育率为71.4％(20/28)。将110枚冻胚移植给7只受体,3只妊娠,产仔12只(5,7),妊娠率为43％(3/7),产仔率为10.9％(12/110),其中囊胚移植1只受体妊娠产仔3只,产仔率为25％(3/12)。

Cryoprezervation of Mouse Embryos by Direct Plunge into Liquid Nitrogen

At the room temperature,292 mouse embryos were incubated respectively in 128. 9 g/L glycerol of PBSSCa phosphate buffer solution with 20% calf serum) and PBSS solution containing 193. 2 g/L glycerol and 85. 6 g/L sucrose for a moment,and then transferred into tube. The tubes with embryos were plunged directly into liquid nitrogen for cryopreservation. After unfreezing,the embryos were washed with PBSS solution containing 342. 3 g/L sucrose to remove the glycerol form them,and then cleaned up with PBSS solution for microscopic examination and evaluation. The embryo recovery rate was 86. 9%(254/292),and the normal morphological rate was 77. 1%(196/254). of them,part of morulae embryos were cultured in vitro,and the development rate of blas-tocysts was 71. 4%(20/28). 110 unfrozen embryos were transferred into 7 recipients,of which 3 became pregnant and 12 young calves were delivered of ( 5, 7) ,the pregnancy rate was 43% (3/7),and the farrowing rate was 10. 9% (12/110). 12 blastocysts were transferred into one recipient wnich became pregnant and produced 3 calves,the farrowing rate was 25//6(3/12). Since this method is both simple and practicable,and there is no need for any expensive equipment and facilities,it is convenient to use in animal production. In addition,the experiment provided some valuable data and experience for the quick freezing of embryos of cattle and other animals.