Journal of Assisted Reproduction and Genetics

, Volume 32, Issue 8, pp 1267–1275 | Cite as

Successful cryopreservation of whole sheep ovary by using DMSO-free cryoprotectant

  • Tianqi Du
  • Lan Chao
  • Shuqin Zhao
  • Linglong Chi
  • Dong Li
  • Yanjun Shen
  • Qing Shi
  • Xiaohui Deng
Fertility Preservation



The study aims to assess the protective effects of dimethyl sulfoxide (DMSO)-free solution based on trehalose on the cryopreservation of a whole sheep ovary and evaluate its use as an efficient cryoprotectant.


Twenty-one ovaries collected from 6- to 8-month-old non-pregnant female sheep were randomly distributed into three groups, namely, a fresh group, a DMSO-free group, and a DMSO group. The morphology, cell apoptosis (by hematoxylin and eosin (HE) staining and terminal deoxynucleotidyltransferase-mediated dUTP nick-end labeling (TUNEL) assay), and mRNA transcript of Bcl-2-associated X protein (BAX) and cold inducible RNA-binding protein (CIRP) (by real-time PCR) of the thawed sheep ovaries and fresh controls were tested to establish a criterion for appraising the results of the cryopreservation.


(i) The histological assessment indicated that the structure of the DMSO-free ovaries remained largely intact and comparable to those of the fresh control groups; whereas, significant damage was observed in the ovaries of the DMSO group (P < 0.05). (ii) The TUNEL assay and mRNA transcript of the BAX assessment showed that the apoptosis parameter in the fresh group was the lowest among all the groups (P < 0.05), and the parameter in the DMSO-free group was significantly lower than that in the DMSO group (P < 0.05). (iii) The level of the CIRP transcripts increased the most in the DMSO-free group followed by the DMSO group and the fresh control group (P < 0.05).


These results indicate that a DMSO-free cryoprotectant solution, especially a trehalose cryoprotectant, is an efficient cryoprotectant and has a beneficial effect on the cryopreservation of whole sheep ovaries.


Whole ovary Sheep Cryopreservation Trehalose Fertility preservation 



The work of this article was done in the Frozen Laboratory of Qilu Hospital of Shandong University.

The authors are grateful to the Shandong Province Science and Technology Research Project (2010GSF10814) and to the National Science Council for financially supporting this study (81370711); the Frozen Laboratory of Qilu Hospital of Shandong University generously provided relevant experimental technology for this study.


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Copyright information

© Springer Science+Business Media New York 2015

Authors and Affiliations

  • Tianqi Du
    • 2
  • Lan Chao
    • 1
  • Shuqin Zhao
    • 3
  • Linglong Chi
    • 4
  • Dong Li
    • 5
  • Yanjun Shen
    • 1
  • Qing Shi
    • 5
  • Xiaohui Deng
    • 1
  1. 1.Center of Reproductive MedicineQilu Hospital of Shandong UniversityJinanPeople’s Republic of China
  2. 2.Center for Reproduction and GeneticsSuzhou Hospital Affiliated to Nanjing Medical UniversitySuzhouChina
  3. 3.Center of Reproductive MedicineZaozhuang Maternal and Children Health HospitalZaozhuangChina
  4. 4.Shandong Qilu Stem Cells Co. LtdJinanChina
  5. 5.Frozen LaboratoryQilu Hospital of Shandong UniversityJinanChina

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