Can frozen-thawed human ovary withstand refreezing-rethawing in the form of cortical strips?
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FERTILITY PRESERVATION
Can frozen-thawed human ovary withstand refreezing-rethawing in the form of cortical strips? Camille Hossay 1 & Alessandra Camboni 1,2 & Luciana Cacciottola 1 & Thu Y. T. Nguyen 1 & Rossella Masciangelo 1 & Jacques Donnez 3 & Marie-Madeleine Dolmans 1,4 Received: 2 June 2020 / Accepted: 29 September 2020 # Springer Science+Business Media, LLC, part of Springer Nature 2020
Abstract Purpose The aim of this study was to elucidate whether ovarian tissue is able to withstand a double freezing-thawing procedure. Methods Human ovarian cortical biopsies from 4 thawed whole ovaries were divided into 4 experimental subgroups: (a) frozenthawed non-grafted group, (b) frozen-thawed xenografted group, (c) refrozen-rethawed non-grafted group, and (d) refrozenrethawed xenografted group. Xenografting was performed using 8 severe combined immunodeficient mice for a total duration of 21 days. The following analyses were conducted: classic hematoxylin and eosin staining, Ki67 immunolabeling, transmission electron microscopy, Masson’s green trichrome, and double CD34 immunostaining. Results Morphologically normal preantral follicles were detected in all groups. We observed a dramatic decline of more than 65% in early preantral follicle survival rates after grafting of both frozen-thawed (p < 0.0001) and refrozen-rethawed (p < 0.0001) ovarian tissue. However, mean follicle densities remained comparable between the frozen-thawed and refrozen-rethawed nongrafted groups, as well as both grafted groups. Equivalent proportions of proliferating early preantral follicles were identified in frozen-thawed and refrozen-rethawed samples, whether the tissue was grafted or not. Furthermore, we did not observe any significant difference in atretic follicle rates between any of the four groups, and the ultrastructural quality of follicles appeared unaffected by the refreezing procedure. Similar proportions of fibrosis were noted in the frozen-thawed and refrozen-rethawed groups, irrespective of grafting. Finally, no significant differences were witnessed in terms of vascularization. Conclusion We were able to demonstrate, for the first time, that refrozen-rethawed ovarian tissue has the same functional characteristics as frozen-thawed ovarian tissue. Keywords Refreezing . Ovarian tissue cryopreservation . Fertility preservation . Xenografting . Electron microscopy
Introduction Over the past two decades, major advances in cancer diagnosis and therapy have resulted in an overall decline in cancer
* Marie-Madeleine Dolmans [email protected] 1
Pôle de Recherche en Gynécologie, Institut de Recherche Expérimentale et Clinique, Université Catholique de Louvain, Avenue Mounier 52, bte. B1.52.02, 1200 Brussels, Belgium
2
Anatomopathology Department, Cliniques Universitaires Saint-Luc, Avenue Hippocrate 10, 1200 Brussels, Belgium
3
Society for Research into Infertility, Avenue Grandchamp 143, 1150 Brussels, Belgium
4
Gynecology Department, Cliniques Universitaires Saint-Luc, Avenue Hippocrate 10, 1200 Brussel
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