Cryopreservation of adult bovine testicular tissue

To develop a practical procedure for cryopreservation of adult bovine testis tissue, the impacts of three cryoprotectans and their concentrations, as well as varying thawing temperatures, on the cell viability of bovine testis tissue after freezing/thawing were examined. The highest testicular cell viabilities came from the media containing dimethyl sulphoxide (DMSO, 85.3±1.2%), propylene glycol (PG, 82±1.0%) and ethylene glycol (EG, 83.4±1.0%) at 10% (V/V) concentration respectively. Using 10% DMSO gave significantly higher spermatogonia percentage (61.1±1.2%, P < 0.001) than processing with 10% PG (54.3±0.6%) or 10% EG (55±1.8%) after differential plating. Thawing in water bath of 37–40°C and 97∼100°C also provided significantly higher viabilities (85.1±1.0 and 85±1.0%, P < 0.001, respectively) and spermatogonia percentages (56.6±2.0 and 56.6±2.6%, P < 0.001, respectively) than that thawing at 4°C (23.4±0.8% for total viability, 8.97 ± 1.0% for spermatogonia percentage). Collectively, 10%DMSO and thawing in 37∼100°C water bath were appropriate for the cryopreservation of bovine testicular tissue and subsequent spermatogonia enrichment.