不同玻璃化冷冻方法对实验兔胚胎超低温保存的影响  

Effect of Different Vitrifications of Rabbits Morula Cryopreservation

作　　者：柴世庆[1] 刘丽均[2] 郁丽丽[2] 柴家前[1] 徐平[2] 

机构地区：[1]山东农业大学动物科技学院,泰安271018 [2]中国科学院上海实验动物中心,上海201615

出　　处：《实验动物与比较医学》2007年第2期90-93,共4页Laboratory Animal and Comparative Medicine

基　　金：国家自然科技资源条件平台项目(2004DKA30650);上海市科技发展基金(0549090007)

摘　　要：目的采用不同的抗冷冻液对实验兔桑椹胚的超低温保存进行探索,以达到经济、有效保存实验兔资源的目的。方法选取三个方案进行实验。方案一:采用改良的EFS40(40%乙二醇+180ppm聚蔗糖(w/v)+0.3mol/L蔗糖PB1溶液)溶液mEFS40作为冷冻保护液,胚胎先在mEFS20溶液中平衡2min,然后移入mEFS40中平衡1min,再移入麦管中,麦管在液氮表面预冷却2min后投入液氮中保存。方案二:将冷冻液和平衡液分别替换为EFS40和EFS20,冷冻步骤同方案一。方案三:使用EFS40溶液作为抗冷冻液和平衡液,胚胎在EFS40中冲洗三遍,移入麦管中后将麦管投入液氮中冷冻保存。冷冻胚胎的复苏均采用25℃水浴加温,然后将胚胎置于0.5mol/L的蔗糖溶液中平衡,再转入PB1溶液中进行培养。结果三个不同方案的冷冻胚胎的复苏率分别为17.1%、69.6%和36.1%。结论采用EFS40作为冷冻保护液和EFS20作为平衡液(方案二)可有效保存实验兔胚胎。Objective EFS40, EFS20, mEFS40, mEFS20 solution was used to cryopreserved rabbits＇ morula economically and effectively. Methods Three protocols were selected in this experiment. Using reEFS40 medium （ modified EFS40） （40% Ethylene glycol＋ 180ppm Ficoll70（W/v）＋ 0.3mol/L Sucrose in PB1） as a cryoprotective solution, rabbit morula were equilibrated two minutes in mEFS20 medium, then equilibrated in mEFS40 cryoprotective solution one minutes. After that they were transferred into plastic insemination straws. The straws were kept on the liquid nitrogen for two minutes. Finally the straws with embryos was put into the liquid nitrogen. The solutions used in project two were replaced with EFS20 and EFS40, the other procedure was the same as the project one. Only EFS40 solution was used in project three, the embryos were washed three times with EFS40 and transferred into the plastic insemination straws, then put the straws into the liquid nitrogen directly. All cryopreserved embryos were thawed in 25℃ water, and incubate in PB1 solution. Results The post-thaw survival in three protocols was 17.1%, 69.6% and 36.1% respectively. Conclusions The rabbits morula can be effectively cryopreserved by using the EFS40 as the cryoprotectant and EFS20 as the equilibrated solution （progeet two）.

关 键 词：冷冻保护液 低温保存 兔 胚胎 

分 类 号：Q813.7[生物学—生物工程]