不同植冰温度下冷冻液的结晶变化及其对小鼠胚胎冷冻的效果

Effects of crystallization of cryoprotectants at different seeding temperature on the developmental ability of mouse embryos

采用程序化冷冻方法 ,对不同植冰温度下几种冷冻液的结晶变化进行了分析 ,并比较了其对小鼠胚胎的冷冻效果。结果表明 ,- 5 .0℃植冰时 ,3种冷冻液 PEF、PGF和 PEGF只有部分结晶 ;当植冰温度为 - 5 .5℃时 ,PEF和 PEGF完全结晶时间为 1 0 min,PGF在 1 5 m in尚未完全结晶 ;- 6 .0℃植冰时 ,3种冷冻液完全结晶时间分别为 8,1 0和 1 0 min;- 6 .5℃植冰后完全结晶时间分别为 5 ,8和 5 min。将小鼠胚胎载入上述 3种冷冻液 ,- 5 .5℃植冰冷冻解冻后 ,囊胚孵化发育率分别为 81 .7% ,6 8.5 %和 87.5 % ,- 6 .0℃植冰时发育率分别为 80 % ,76 .5 %和 90 .9% ,- 6 .5℃植冰后的发育率分别为 1 0 % ,1 3.3%和 38.4 %。据此得出 ,在 - 5 .5或 - 6 .0℃时植冰 ,以

Crystallization changes of various cryoprotectants sections were analysized at different seeding temperature using programmed freezing procedure.When seeded at -5.0 ℃,PBS was freezed completely within 2 min,PBS+ethylene glycol (PEF),PBS+glycerol (PGF) and PBS+ethylene glycol+glycerol (PEGF) were not crystallized entirely although the process lasted for 15 min;When seeding temperature was -5.5 ℃,PEF and PEGF were crystallized entirely in 10 min,but PGF was not in 15 min;While seeding temperature reduced at -6.0 ℃ and -6.5 ℃,the three kinds of cryoprotectants were solidified in 8 min,10 min and 10 min vs 5 min,8 min and 5 min respectively.Mouse embryos were exposed to the three cryopreservative compounds and seeded at -5.5 ℃ and -6.0 ℃ followed lauching into liquid nitrogen at -35 ℃,after thawing and in vitro culture,percentage of hatching embryos were 81.7%,68.5% and 87.5% vs 80%,76.5% and 90.9% ,they both had higher developmental abilities than that seeded at -6.5℃ (10%,13.3% and 38.4%).Compared with PEF and PGF,PEGF had better effect on the freezing of mice embryos.