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Whole sheep ovary cryopreservation: evaluation of a slow freezing protocol with dimethylsulphoxide

  • FERTILITY PRESERVATION
  • Published:
Journal of Assisted Reproduction and Genetics Aims and scope Submit manuscript

Abstract

Purpose

To evaluate a slow freezing method for whole ovary cryopreservation by evaluating effects of added cryoprotectant.

Methods

Sheep ovaries were isolated during surgery, flushed with either Ringer-Acetate or dimethylsulphoxide and cryopreserved by slow freezing. After rapid thawing, viability was assessed by ovarian in vitro perfusion, cell culture, histology and fluorescent live-dead assay.

Results

Production of cyclic AMP and progesterone was slightly higher in the dimethylsulphoxide group. Cultured ovarian cells from dimethylsulphoxide-preserved ovaries secreted larger amounts of progesterone than cells from Ringer-Acetate preserved. Light microscopy of ovarian biopsies obtained after perfusion, revealed well-preserved tissue in the dimethysulphoxide group but not in the Ringer-Acetate group. The density of small follicles and ovarian cell viability were higher in dimethysulphoxide ovaries compared to Ringer-Acetate ovaries.

Conclusions

Equilibrium with its protective effect can be achieved by slow freezing protocol, with an additional protective effect by the presence of dimethylsulphoxide.

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Acknowledgments

Financial support: Supported by grants from: Swedish Research Council, Stockholm, Sweden; Sahlgrenska Academy ALF, Göteborg, Sweden; Hjalmar Svensson’s Research Foundation, Göteborg, Sweden; and Assar Gabrielsson’s Research Foundation, Göteborg, Sweden.

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Authors and Affiliations

  1. Department of Obstetrics and Gynecology, Sahlgrenska Academy, University of Gothenburg, SE-41345, Göteborg, Sweden

    Milan Milenkovic, Ann Wallin & Mats Brännström

  2. Department of Obstetrics and Gynecology, University of British Columbia, Vancouver, BC, Canada

    Manda Ghahremani

Authors
  1. Milan Milenkovic
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  2. Ann Wallin
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  3. Manda Ghahremani
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  4. Mats Brännström
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Corresponding author

Correspondence to Milan Milenkovic.

Additional information

Capsule The study evaluates the impact of dimethylsulphoxide as a cryoprotectant for whole ovary cryopreservation.

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Milenkovic, M., Wallin, A., Ghahremani, M. et al. Whole sheep ovary cryopreservation: evaluation of a slow freezing protocol with dimethylsulphoxide. J Assist Reprod Genet 28, 7–14 (2011). https://doi.org/10.1007/s10815-010-9477-5

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  • DOI: https://doi.org/10.1007/s10815-010-9477-5

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