两种玻璃化胚胎冷冻方法对不同品系小鼠胚胎冷冻效果比较

摘要

Objective To compare the effect of two vitrification-cryopreservation methods of embryos of several mouse strains.Methods The two-cell embryos from six mouse strains (KM,ICR,BALB/c,C57BL/6J,OB/OB,LAP/ rtTA0F59) were cryopreserved and thawed,using EFS and DAP cryoprotectants,respectively.The recovery rate in vitro and implantation rate in vivo were determined to evaluate the efficacy of the two methods.Results The average post-thaw survival rates of the embryos of different strains preserved in EFS were 69.97％ (47.9％ to 83.6％),and preserved in DAP 47.23％ (26.3％ to 76.7％).After 2-cell embryos were transferred to the oviducts of pseudopregnant recipient in all the strains,the implantation rates of EFS-vitrified embryos were ranged from 1.75％ to 45.0％ (average 27.23％) and of DAP-vitrified embryos from 7.0％ to 46.3％ (average 31.43％).With the respect of recovery rate,EFS was preferred to DAP.No significant differences (P ＞ 0.05) were found between the implantation rates of EFS and DAP in KM and ICR strains.But the implantation rates of BALB/c,C57BL/6J,OB/OB,and LAP/rtTA0F59 mouse embryos in EFS were higher than those in DAP,respectively.Conclusions EFS cryoprotectant is suitable for both the KM and ICR mouse embryos vitrification-cryopreservation.DAP is somehow better than EFS for cryopreservation of C57BL/6J,OB/OB and LAP/rtTA0F59 mouse embryos,but with a non-significant difference between the two cryoprotectants.Both EFS and DAP show a rather low implantation rate of the BALB/c mouse embryos,therefore,further studies are needed to improve this issue.%目的 探讨EFS和DAP两种玻璃化冷冻方法对不同品系小鼠胚胎冷冻的效果.方法 6个品系小鼠(KM、ICR、BALB/c、C57BL/6J、OB/OB、LAP/aTA0F59)的2-cell胚胎分别用EFS和DAP两种玻璃化冷冻方法进行冷冻和复苏,比较两种冷冻方法的胚胎复苏率和着床率.结果 6个品系小鼠冷冻胚胎EFS方法的平均复苏率为69.97％ (47.9％～83.6％),DAP方法的平均复苏率47.23％ (26.3％ ～76.7％),EFS方法明显优于DAP方法.其中KM、ICR和BALB/c小鼠EFS方法的冷冻复苏率显著高于DAP方法(P＜0.01)；冻融胚胎移植后EFS方法的平均着床率27.23％(1.75％ ～45.0％),DAP方法的平均着床率31.43％(7.0％～46.3％).除KM、ICR小鼠外,其他4个品系小鼠的着床率DAP方法高于EFS方法.结论 KM和ICR远交群小鼠胚胎适合用EFS方法冷冻保存；C57BL/6J、OB/OB、LAP/rtTA0F59三个品系小鼠DAP方法优于EFS方法,但差异不大；BALB/c小鼠两种玻璃化冷冻方法的冻融胚胎着床率均较低,需进一步研究.