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Damage to fetal bovine ovarian tissue caused by cryoprotectant exposure and vitrification is mitigated during tissue culture

  • Fertility Preservation
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Abstract

Purpose

The objective of this study is to characterize the impact of exposure to cryoprotectants followed by vitrification on primordial follicle survival and activation using a fetal bovine model.

Methods

In the first study, fetal bovine cortical pieces were exposed to cryoprotectants with or without sucrose and cultured up to 7 days in the presence or absence of insulin. In the second study, cortical pieces were exposed to cryoprotectants with or without sucrose, vitrified, and cultured up to 7 days after warming in the presence or absence of insulin. Viability and morphology of follicles, as well as proliferation and/or DNA repair in ovarian tissue were analyzed.

Results

When compared to non-exposed controls, normal follicular morphology was affected in groups exposed to cryoprotectants only immediately post-exposure and after 1 day of culture, but improved by day 3 and did not significantly differ by day 7. Similarly, normal follicular morphology was compromised in vitrified groups after warming and on day 1 compared to controls, but improved by days 3 and 7. Proliferation and/or DNA repair in ovarian tissue was not affected by vitrification in this model. Cryoprotectant exposure and vitrification of ovarian tissue did not impair the activation of primordial follicles in response to insulin, although activation was delayed relative to non-exposed controls. Interestingly, sucrose had no noticeable protective effect.

Conclusion

Vitrified fetal bovine ovarian tissue has the intrinsic capacity to mitigate the immediate damage to primordial follicles’ morphology and retains the capacity to activate. These findings provide a basis for a successful cryopreservation protocol for ovarian cortical tissue in other species including humans.

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Acknowledgments

The authors thank Cargill Regional Beef for the donation of bovine ovaries. The cooperation of John Couture at Cargill is gratefully acknowledged. The authors also thank Dr. Mark Roberson for the use of his storage facilities and Mary Lou Norman for her assistance with histological preparations.

Conflict of interest

The authors declare they have no competing interests.

Funding

This research was supported by the United States Department of Agriculture Multistate Project (NE-1227).

Author information

Authors and Affiliations

  1. Smithsonian Conservation Biology Institute, National Zoological Park, Washington, DC, 20008, USA

    Lara Mouttham & Pierre Comizzoli

  2. Department of Biomedical Sciences, Cornell University, Ithaca, NY, USA

    Lara Mouttham & Joanne E. Fortune

Authors
  1. Lara Mouttham
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  2. Joanne E. Fortune
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  3. Pierre Comizzoli
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Correspondence to Pierre Comizzoli.

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Capsule

Cryoprotectant exposure and vitrification of fetal bovine ovarian tissue do not cause long-term damage to follicle structure or affect the capacity of primordial follicles to activate.

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Mouttham, L., Fortune, J.E. & Comizzoli, P. Damage to fetal bovine ovarian tissue caused by cryoprotectant exposure and vitrification is mitigated during tissue culture. J Assist Reprod Genet 32, 1239–1250 (2015). https://doi.org/10.1007/s10815-015-0543-x

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  • DOI: https://doi.org/10.1007/s10815-015-0543-x

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