摘要
本试验采用高浓度乙二醇 ( EG-8)作为胚胎冷冻保护剂 ,以高浓度的半乳糖作为解冻稀释液 ,对 MMTV-Wnt-1转基因小鼠的早期囊胚进行玻璃化冷冻保存。结果在 EG-2中平衡 4~ 6min和解冻时间为 3~ 5min时获得了最佳冷冻效果。此条件下冷冻胚胎的总回收率为 88.61 % ( 1 0 9/ 1 2 3 ) ;回收胚胎在体外发育率达 88.99% ( 97/ 1 0 9) ;孵化率达81 .65% ( 89/ 1 0 9) ;发育胚胎的移植出生率为 3 8.3 6% ( 69/ 1 81 ) ;冷冻胚胎移植出生后经PCR检测一半小鼠整合阳性。
The high concentration ethylene glycol (EG-8, 8 mol/L) and galactose(1.7 mol/L) were used as a cryoprotectant and thawing diluents for embryo cryopreservation by vitrification in MMTV-Wnt-1 transgenic mice. The best results of cryopreservation were obtained if those embryos kept balance in EG-2 (2mol/L) for 4~6 minute before freeze and thawing 3~5 minute in galactose. The percentage of embryos cryopreserved and retrieve is 88.61%(109/123); there is 88.99%(97/109) of these embryos develop and 81.65%(89/109) developing to hatch blastocysts. After these development embryos were transfer to pseudocyesis mouse, 38.36%(69/181) developed to live young, and a half of these is positive mouse on the PCR test. It follows that high concentration ethylene glycol and galactose as a cryoprotectant for cryopreseveration by vitrification is effective.
出处
《上海实验动物科学》
2001年第4期205-208,共4页
Shanghai Laboratory Animal Science
关键词
乙二醇
转基因小鼠
胚胎
冷冻保存
Ethylene glycol
Transgenic mice
Embryo cryopreservation