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Updated: Jun 8, 2026
Vitrification of In Vitro Matured Oocytes Collected from Adult and Prepubertal Ovaries in Sheep
Published on: July 10, 2021
Whole sheep ovary cryopreservation: evaluation of a slow freezing protocol with dimethylsulphoxide
Milan Milenkovic1, Ann Wallin, Manda Ghahremani
1Department of Obstetrics and Gynecology, Sahlgrenska Academy, University of Gothenburg, SE-41345, Göteborg, Sweden. milan.milenkovic@obgyn.gu.se
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.
