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A modified cryoloop vitrification protocol in the cryopreservation of mature mouse oocytes

Published online by Cambridge University Press:  01 August 2009

Zengyan Wang ,
Zhengyi Sun ,
Ying Chen  and
Fangfang He
Zengyan Wang
Affiliation:
Department of Gynecology and Obstetrics, Peking Union Medical College Hospital, Chinese Academy of Medical Science, Beijing, China.
Zhengyi Sun
Affiliation:
Department of Gynecology and Obstetrics, Peking Union Medical College Hospital, Chinese Academy of Medical Science, Beijing, China.
Ying Chen
Affiliation:
Department of Gynecology and Obstetrics, Peking Union Medical College Hospital, Chinese Academy of Medical Science, Beijing, China.
Fangfang He*
Affiliation:
Department of Gynecology and Obstetrics, Peking Union Medical College Hospital, 1 Shuaifuyuan Alley, Dongcheng District, Beijing, China. Department of Gynecology and Obstetrics, Peking Union Medical College Hospital, Chinese Academy of Medical Science, Beijing, China.
*
All correspondence to Fangfang He, Department of Gynecology and Obstetrics, Peking Union Medical College Hospital, 1 Shuaifuyuan Alley, Dongcheng District, Beijing, China. Tel: +610 65296223. Fax: +8610 65296223. e-mail: hefangfangmd@yahoo.com.cn
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Summary

In this study, we examined a modified cryoloop vitrification protocol in the cryopreservation of mature mouse oocytes. The mature mouse oocytes were first vitrified and then warmed up in a modified cryoloop vitrification medium [15% ethylene glycol (EG) + 15% dimethyl sulphoxide (ME2SO) + 5.8 mg/ml Ficoll 400 (F) + 0.58 mol/l sucrose (S)]. These oocytes were later studied along with fresh oocytes, which served as the control group.

Based on the post-warm-up incubation time, the oocytes in the study group were divided into three subgroups: 0 h, 1 h and 2 h. We then examined the configurations of spindles and chromosomes, the fragmentation of DNA, and the oocyte's ability to be fertilized and developed into blastocysts. By evaluating the vitrified oocytes' morphology, we confirmed that 601 out of 612 (98.2%) oocytes survived this protocol. The percentage of oocytes with normal spindle and chromosome configurations in the study groups 0 h, 1 h and 2 h were all quite similar to each other and not statistically different from that of the control group. Similar results were also observed in the percentage of oocytes containing fragmented DNA. The fertilization rate and blastocyst formation rate of the thawed oocytes were not statistically different from that of the control group either. However, if not handled properly (too much remnant medium on oocytes in the process of freezing or too long a time of oocytes in the vitrification medium before freezing), the cryopreserved oocytes could show dramatic difference from the control group in terms of the morphologically survival rate, the configuration of the spindles and chromosomes, and the DNA fragmentation. In conclusion, when followed correctly, this modified cryoloop vitrification protocol had little effect on the survival rate and development potential of mature mouse oocytes.

Keywords

ChromosomeDNA fragmentationOocytesSpindleVitrification
Type
Research Article
Information
Zygote , Volume 17 , Issue 3 , August 2009 , pp. 217 - 224
Copyright
Copyright © Cambridge University Press 2009

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