Effects of vitrifying mouse embryos from different periods and different sources
LIU Yun1, HU Jie2, CHEN Xiao-fang1, YIN Zhao-hui1, MI Wu-xuan1, HU Jia-bo2
（1.Human Reproduction Medicine Center, the Fourth Affiliated Hospital of Jiangsu University, Zhenjiang Jiangsu 212001; 2.School of Medical Science and Laboratory Medicine, Jiangsu University, Zhenjiang Jiangsu 212013, China）
Abstract：Objective: To compare the postthaw survival rate and the success rate in continuing to grow to blastocyst of vitrified mouse embryos from different periods and different sources. Methods: The 2-cell, 4-cell, 8-cell period mouse embryos were obtained through fertilization in vitro and in vivo respectively. These embryos were vitrified and thawed. We got the rate of their survival and blastocyst formation. And, we took those fresh embryos as the control group. Results: The survival rate of blastocyst formation of post-thaw 2-cell, 4-cell and 8-cell period embryos from fertilization in vitro and in vivo were 70.67%/65.33% and 72.06%/67.62%, 88.00%/81.33% and 89.71%/83.82% and 93.33%/92.00% and 94.12%/94.12% respectively. The blastocyst formation rate in the phrase of 2-cell and 4-cell frozen embryos was lower than the control group (P＜0.05).No significant difference was found between the group of 8-cell frozen embryos and the control group. In addition, fertilization modes had no significant effect on the survival rate and the blastocyst formation rate(P>0.05). Conclusion: Vitrification technology is an effective method in preserving mouse embryos. And the 8-cell period embryos might have the best cryopreservation effect.
收稿日期: 2013-01-08
基金资助:
镇江市科技基金资助项目(FZ2011057)；江苏大学科研立项资助项目(11A394)
通讯作者:
胡嘉波（通讯作者），副教授，硕士生导师，Email: hu@ujs.edu.cn
作者简介: 刘云（1969—），女，江苏扬中人，副主任技师，主要从事生殖医学实验研究;
引用本文:
刘云，胡杰，陈小芳，殷朝晖，糜戊煊，胡嘉波. 不同时期和不同来源鼠胚玻璃化冷冻效果的比较[J]. 江苏大学学报:医学版, 2013, 23(2): 120-123,129.
LIU Yun1, HU Jie2, CHEN Xiao-fang1, YIN Zhao-hui1, MI Wu-xuan1, HU Jia-bo2. Effects of vitrifying mouse embryos from different periods and different sources. Journal of Jiangsu University(Medicine Edition), 2013, 23(2): 120-123,129.
[1]Wong JY, Wong AY. Phasing-in of vitrification into routine practice: why, how, and what[J]. Hong Kong Med J, 2011,17(2):119-126.[2]Abdelhafez F, Xu J, Goldberg J, et al. Vitrification in open and closed carriers at different cell stages: assessment of embryo survival, development, DNA integrity and stability during vapor phase storage for transport[J]. BMC Biotechnol, 2011,11:29.[3]Andras N,Marina G,Kristina V,等.小鼠胚胎实验操作手册[M].孙青原，陈大元译.3版.北京：化学工业出版社，2006：156-158,420-422.[4]鲁栋梁，崔曙，陈在贤. EDS-40 玻璃化液冻存小鼠4 细胞胚胎的研究[J]. 四川医学,2012, 33(2):198-200.[5]Whittingham DG, Leibo SP, Mazur P. Survival of mouse embryos frozen to-196 degrees and269 degrees C[J]. Science, 1972, 178(4059): 411-414.[6]Wilmut I. The effect of cooling rate, warming rate,cryoprotective agent and stage of development on survival of mouse embryos during freezing and thawing[J]. Life Sci, 1972, 11(22): 1071-1079.[7]Rall WF，Fahy GM. Ice-free cryopreservation of mouse embryos at-196 degrees C by vitrification[J]. Nature, 1985, 313(6003):573-575.[8]Mukaida T, Wada S, Takahashi K, et al. Vitrification of human embryos based on the assessment of suitable conditions for 8-cell mouse embryos[J].Hum Reprod, 1998,13(10):2874-2879.[9]陈莉，刘琦．玻璃化冷冻技术在人类辅助生殖中的应用\[J\].中华男科学杂志，2010,16(6): 556-560.[10]赵丹凤，刘丽均，赵立虎，等．F1代和ICR小鼠体外、体内受精后8细胞胚胎玻璃化冷冻的比较[J]．现代畜牧兽医,2006(12):10-12.[11]连安，曲玉秀，黄裕香，等．小鼠不同阶段胚胎玻璃化冷冻保存的比较研究[J].实验生物学报，2003，36（4）：279-282.[12]Cseh S, Corselli J, Nehlsen-Cannarella SL, et al. The effect of quick-freezing in ethylene glycol on morphological in vitro development of mouse embryos frozen at different preimplantation stages[J].Theriogenology, 1997, 48(1): 43-50.[13]高珊，张建军, 薛孝先，等.胚胎发育期和冷冻保护剂剂量对Nakagata小鼠胚胎冷冻效果的影响[J]. 中国比较医学杂志, 2007,17(11):666-669.[14]Vajta G, Holm P, Kuwayama M, et al. Open Pulled Straw(OPS) vitrification:a new way to reduce cryoinjuries of bovine ova and embryos[J]. Mol Reprod Dev,1998,51(1): 53-58.
2013, Vol. 23 
