Whole sheep ovary cryopreservation: evaluation of a slow freezing protocol with dimethylsulphoxide.
OBJECTIVE
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.
Milenkovic
Milan
M
Department of Obstetrics and Gynecology, Sahlgrenska Academy, University of Gothenburg, SE-41345, Göteborg, Sweden. milan.milenkovic@obgyn.gu.se
Wallin
Ann
A
Ghahremani
Manda
M
Brännström
Mats
M
D000818:Animals; D002470:Cell Survival; D002478:Cells, Cultured; D015925:Cryopreservation / Q000379:methods* / Q000662:veterinary*; D003451:Cryoprotective Agents / Q000494:pharmacology*; D000242:Cyclic AMP / Q000378:metabolism; D004121:Dimethyl Sulfoxide / Q000494:pharmacology*; D005260:Female; D005615:Freezing; D007552:Isotonic Solutions / Q000494:pharmacology; D006080:Ovarian Follicle / Q000166:cytology / Q000637:transplantation; D010477:Perfusion; D011374:Progesterone / Q000378:metabolism; D034561:Sheep, Domestic
D003451:Cryoprotective Agents; D007552:Isotonic Solutions; C028570:Ringer's acetate; D011374:Progesterone; D000242:Cyclic AMP; D004121:Dimethyl Sulfoxide