不同冷冻保护剂和冷冻方法对小鼠胚胎发育的影响

Effects of different cryoprouectants and cryopreservation protocols on the development of mouse 2-cell embryos

目的评价不同冷冻保护剂和不同冷冻方法对小鼠2-细胞胚胎发育的影响。方法采用丙二醇(PROH)、乙烯乙二醇(EG)、二甲基亚砜(DMSO)和甘油(G)4种不同冷冻保护剂,慢速冷冻、Vit-Master玻璃化冷冻两种冷冻方式冷冻小鼠2-细胞胚胎,观察胚胎的成活率。结果采用慢速冷冻方式冷冻小鼠2-细胞胚胎时,以PROH为冷冻保护剂的胚胎成活率、囊胚形成率、囊胚孵出率显著高于以DMSO、G为冷冻保护剂的结果(P<0.05),而以DMSO、G、EG为冷冻保护剂的结果比较差异无显著性意义(P>0.05);采用Vit-Master玻璃化冷冻方式冷冻小鼠2-细胞胚胎时,以EG为冷冻保护剂的胚胎成活率、囊胚形成率、囊胚孵出率显著高于以PROH、DMSO、G为冷冻保护剂的结果(P<0.05),而以PROH、DMSO、G为冷冻保护剂的结果比较差异无显著性意义(P>0.05)。结论PROH是慢速冷冻小鼠2-细胞胚胎理想的冷冻保护剂;EG是Vit-Master玻璃化冷冻小鼠2-细胞理想的冷冻保护剂,有可能替代慢速冷冻法。

Objective To evaluate the effects of different cryoprotectants and different cryopreservation protocols on the development of mouse 2-cell embryos. Methods Mouse 2-cell embryos were cryopreserved using propylene glycol （ PROH） , ethylene glycol （ EG）, demethyl-sulphoxide （DMSO） or glycerol （G） as cryoprotectants with slow-freezing or Vit-Master vitrifi- cation protocol. After thawing, the survival rate, blastocyst formation rate and blastocyst hatching rate of the embryos were compared with each other. Results When the mouse 2-cell embryos were cryopreserved by the slow-freezing, the survival rate, the blastocyst formation rate and the blastocyst hatching rate of the embryos with PROH were significantly higher than those with DMSO and G [ P 〈0. 05）, but not significantly different among those with DMSO, G or EG [P 〉 0. 05）. When the mouse 2-cell embryos were cryopreserved by Vit-Master vitrification, the survival rate, the blastocyst formation rate and the blastocyst hatching rate of the embryos with EG were significantly higher than those with PROH, DMSO or G （P 〈 0. 05 ）. However, there were no significant differences among those with PROH, DMSO or G （ P 〉 0. 05 ）. Conclusion PROH is an optimal cryoprotectant for the cryopreservation of mouse 2-cell embryos when slow-freezing protocol is used. EG is an optimal cryoprotectant for the cryopresevation of mouse 2-cell embryos when Vit-Master vitrification protocol is adopted, which may substitute for slow-freezing protocol that has been commonly used in clinical and laboratory practice.