Effects of Cryoprotectant Agents on Equine Ovarian Biopsy Fragments in Preparation for Cryopreservation

Abstract The exposure effect of cryoprotectant agents (CPAs) on morphology of preantral follicles (PAFs), stromal cell and PAF densities, and area of equine ovarian fragments were evaluated. Three independent experiments with identical methodologies were performed. Each experiment was composed of one CPA (dimethyl sulfoxide, ethylene glycol, or propylene glycol) and was performed in three replicates. Ovarian biopsy fragments were harvested from six mares in each experiment and submitted to the cryoprotectants using four times of exposure (0, 10, 15, and 20 minutes). PAF and stromal cell densities, and area of the fragments were not affected ( P > .05) by any of the CPAs throughout the time of exposure. However, the morphology of the PAFs was affected ( P P P > .05) throughout the times of exposure. Positive correlations ( r = 0.57–0.77; P