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

  • Fertility Preservation
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Abstract

Purpose

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.

Method

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.

Results

(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).

Conclusions

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.

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Acknowledgments

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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    Authors and Affiliations

    1. Center of Reproductive Medicine, Qilu Hospital of Shandong University, 107 West Wenhua Road, Jinan, Shandong Province, 250012, People’s Republic of China

      Lan Chao, Yanjun Shen & Xiaohui Deng

    2. Center for Reproduction and Genetics, Suzhou Hospital Affiliated to Nanjing Medical University, 26 Daoqian Street, Suzhou, Jiangsu, 215002, China

      Tianqi Du

    3. Center of Reproductive Medicine, Zaozhuang Maternal and Children Health Hospital, Zaozhuang, 370400, China

      Shuqin Zhao

    4. Shandong Qilu Stem Cells Co. Ltd, Jinan, 250000, China

      Linglong Chi

    5. Frozen Laboratory, Qilu Hospital of Shandong University, Jinan, 250012, China

      Dong Li & Qing Shi

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    1. Tianqi Du
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    Correspondence to Xiaohui Deng.

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    Capsule

    These results indicate that a DMSO-free and trehalose-containing cryoprotectant solution has a beneficial effect on the cryopreservation of intact sheep ovaries.

    Tianqi Du and Lan Chao contributed equally to this work.

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    Du, T., Chao, L., Zhao, S. et al. Successful cryopreservation of whole sheep ovary by using DMSO-free cryoprotectant. J Assist Reprod Genet 32, 1267–1275 (2015). https://doi.org/10.1007/s10815-015-0513-3

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    • DOI: https://doi.org/10.1007/s10815-015-0513-3

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