Abstract
Purpose
The aim of this study was to determine the best combination in terms of cryopreservation techniques and vascular bed preparation before grafting in order to obtain functional ovarian tissue after transplantation.
Methods
Five cynomolgus monkeys were used. Strips from 10 ovaries were cryopreserved, 5 by vitrification (V), and 5 by slow-freezing (SF). Pieces of fresh ovarian tissue were used for controls. After 1 month, the strips were autografted to two different vascular beds, healed (HB) or freshly decorticated (FDB), constituting four study groups: SF-HB, SF-FDB, V-HB, and V-FDB. These were compared to fresh tissue. After 6 months, the ovaries were removed and several parameters analyzed: follicle quality, stage, density, proliferation, apoptosis, functionality, vascularization, and fibrosis. Mixed effect linear regression models were built to assess the impact of cryopreservation and vascular bed preparation on ovarian tissue viability and functionality. p values were adjusted for multiple testing using the Benjamini-Hochberg method, and q values < 0.20 were considered significant in order to achieve a 20 % false discovery rate.
Results
Compared to fresh tissue, no difference was observed in the percentage of morphologically normal follicles, while a significant increase was noted in the follicle proliferation rate (41 %, q = 0.19), percentage of antral follicles (12 %, q = 0.14), and number of vessels per area (3.3 times, q = 0.07) in the V-FDB group.
Conclusions
Vitrification associated with FDB vascular bed preparation is the best combination to obtain functional autografted ovarian tissue. Further studies are nevertheless required, with confirmed pregnancies and live births before introducing the procedure into clinical practice.
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Acknowledgments
The authors thank Maria Dolores Gonzalez and Olivier Van Kerk for their technical support and Mira Hryniuk for revising the English language of the manuscript. This work was supported by grants from the Fonds National de la Recherche Scientifique de Belgique (5/4/150/5, W.0056.15 and T.0077.14), Fonds Spéciaux de Recherche, Fondation Saint Luc, Foundation Against Cancer, and donations from Mr. Pietro Ferrero, Baron Frère, and Viscount Philippe de Spoelberch.
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Ethical approval
Approval for this study was obtained from the Animal Ethics Review Board of the Université Catholique de Louvain (2011/UCL/MD/012). All procedures were conducted according to guidelines established by the abovementioned committee and according to Belgian (AR 29, May 2013) and European (Directive 2010/63/EU) legislation on the care and use of laboratory animals. This manuscript does not involve any studies with human participants performed by any of the authors.
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The authors declare that they have no conflict of interests.
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Capsule Vitrification associated with freshly decorticated vascular bed preparation appears to be the best combination to obtain functional autografted ovarian tissue.
M. M. Dolmans and M. M. Binda contributed equally to this work.
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Dolmans, M.M., Binda, M.M., Jacobs, S. et al. Impact of the cryopreservation technique and vascular bed on ovarian tissue transplantation in cynomolgus monkeys. J Assist Reprod Genet 32, 1251–1262 (2015). https://doi.org/10.1007/s10815-015-0542-y
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DOI: https://doi.org/10.1007/s10815-015-0542-y