Complete protection against cryodamage of cryopreserved whole bovine and human ovaries using DMSO as a cryoprotectant

Johan R Westphal,Ronald Peek,Renne Gerritse,Didi D M Braat,Catharina C M Beerendonk

doi:10.1007/s10815-017-0963-x

Abstract

PurposeThis study aims to determine the optimal cryopreservation protocol for whole ovaries intended for preservation of fertility in women.MethodsWe investigated the optimal cryopreservation procedure for whole ovaries in a bovine model. The following parameters were investigated to determine their effect on ovarian tissue viability: type of cryoprotectant, administration route of the cryoprotectant (perfusion and/or submersion), and the maximum tolerable interval between death of the animal and start of the cryopreservation process. The resulting optimal cryopreservation procedure for bovine ovaries was subsequently tested on human ovaries. In vitro glucose uptake, histology, and immunohistochemistry were used to assess the integrity of the ovarian tissue.ResultsStarting the cryopreservation procedure (including perfusion with and submersion in DMSO) within 10–15 min after death of the animal proved critical, resulting in a 90–100% protection level against cryodamage. When cryopreserving human ovaries using the same protocol, over 95% protection against cryodamage was observed on all tissue levels. In addition, no apparent morphological damage to either the follicles or the vascular endothelium was observed.ConclusionOur findings suggest that using the optimized protocol presented in this paper allows good cryopreservation of whole human ovaries and represents an important step in considering whole ovary autotransplantation for clinically applied fertility preservation.

Highlights

With treatment modalities improving over the years, the number of cancer patients that survive their disease is increasing
We have shown that submerging an intact bovine ovary in a 10% dimethyl sulfoxide (DMSO) solution for 15 min fully protected the cortical layer of the ovary, while leaving deeper tissue layers unprotected against cryodamage [26]
We found it was crucial to confine the period of warm ischemia to an absolute minimum

Summary

Introduction

With treatment modalities improving over the years, the number of cancer patients that survive their disease is increasing Parallel to this encouraging development, growing attention is being given to the quality of life after cure, including the ability to start a family. Cryopreservation of cortical ovarian tissue strips is one option for fertility preservation These tissue strips can, after the patient has been cured of her disease, be thawed and autotransplanted to restore fertility. To date, this option for fertility preservation has already led to the birth of at least 86 children worldwide, indicating the viability of this option [6,7,8]

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