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使用不同冷冻保护剂对小鼠胚胎进行慢速冷冻和玻璃化冷冻的比较研究。

Comparative study between slow freezing and vitrification of mouse embryos using different cryoprotectants.

作者信息

Liu W-X, Luo M-J, Huang P, Yue L-M, Wang L, Zhao C-Y, He Y-P, Zhang J-H, Zheng Y

机构信息

Department of Physiology, West China School of Preclinical and Forensic Medicine, Sichuan University, Chengdu, China.

出版信息

Reprod Domest Anim. 2009 Oct;44(5):788-91. doi: 10.1111/j.1439-0531.2008.01078.x. Epub 2008 Oct 30.

Abstract

The objective of this study was to evaluate the effects of different cryoprotectants and different cryopreservation protocols on the development of mouse eight-cell embryos. Mouse eight-cell embryos were cryopreserved by using propylene glycerol (PROH), ethylene glycerol (EG), dimethyl sulfoxide (DMSO) or glycerol (G) as cryoprotectant with slow-freezing or Vit-Master vitrification protocol. After thawing, the survival rate, blastocyst formation rate and blastocyst hatching rate of the embryos were compared. When the mouse eight-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 of DMSO and G (p < 0.05, respectively), but not significantly different among those of DMSO, G and EG (p > 0.05, respectively), and not significantly different between those of PROH and EG (p > 0.05, respectively). When the mouse eight-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 of PROH, DMSO and G (p < 0.05, respectively). Yet, there were no significant differences among those of PROH, DMSO and G (p > 0.05, respectively). In conclusion, PROH was the optimal cryoprotectant for the cryopreservation of mouse eight-cell embryos by slow-freezing protocol. EG was the optimal cryoprotectant for the cryopresevation of mouse eight-cell embryos by Vit-Master vitrification protocol, which may be commonly used in clinical and laboratory practice.

摘要

本研究的目的是评估不同冷冻保护剂和不同冷冻保存方案对小鼠八细胞胚胎发育的影响。以丙二醇(PROH)、乙二醇(EG)、二甲基亚砜(DMSO)或甘油(G)作为冷冻保护剂,采用慢速冷冻或Vit-Master玻璃化方案对小鼠八细胞胚胎进行冷冻保存。解冻后,比较胚胎的存活率、囊胚形成率和囊胚孵化率。当采用慢速冷冻法冷冻保存小鼠八细胞胚胎时,使用PROH的胚胎的存活率、囊胚形成率和囊胚孵化率显著高于使用DMSO和G的胚胎(p分别<0.05),但DMSO、G和EG之间无显著差异(p分别>0.05),PROH和EG之间也无显著差异(p分别>0.05)。当采用Vit-Master玻璃化法冷冻保存小鼠八细胞胚胎时,使用EG的胚胎的存活率、囊胚形成率和囊胚孵化率显著高于使用PROH、DMSO和G的胚胎(p分别<0.05)。然而,PROH、DMSO和G之间无显著差异(p分别>0.05)。总之,PROH是采用慢速冷冻方案冷冻保存小鼠八细胞胚胎的最佳冷冻保护剂。EG是采用Vit-Master玻璃化方案冷冻保存小鼠八细胞胚胎的最佳冷冻保护剂,这可能在临床和实验室实践中普遍应用。

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