Influence of Culture Media on the Motility and Viability of Cryopreserved Stallion Spermatozoa

Thawed cryopreserved stallion semen was washed in PBS+0.3%BSA at 900 g for 5¢and cultured in 35 mm petri dishes in two different media: SOF+0.3%BSA (SOF) and TCM199+0.3% BSA (TCM) supplemented or not with 1500 UI/ml SOD. At 0, 3, 6, 9, 24 and 48 h of culture motility, viability and acrosome status were evaluated with propidium iodide and FITC-PSA. Results indicate that mass motility, measured as a 0–10 grade scale by three different operators, was higher in SOF than in TCM (7.3 vs. 3.7, 6 vs. 3, 4.3 vs. 2.3, 2 vs. 1, 0.7 vs. 0 at 3, 6, 9, 12, 24 and 48 h of culture respectively). SOD supplementation increased spermatozoa motility compared to the basic media (SOF: 8.7, 8.3, 6.3, 3, 2.39; TCM: 5.3, 4, 2.7,1, 0 for the same times). Cell viability was higher (p < 0:01) in SOF compared to TCM (3h:50.0 and 45, 6h : 54.8% vs. 32.3%, 9h : 39.1% vs. 15.5%, 24h : 14.5% vs. 2.0%, 48h : 5.5% vs. 0 for SOF and TCM, respectively). The viability rate was increased (p < 0:05) in TCM+SOD (3h : 45.1 vs. 54.1, 6h : 32.3 vs. 44.1, 9h : 15.5 vs. 37.5 and 24h : 2 vs. 10) but not in SOF+SOD. The rate of live spermatozoa with intact acrosome membrane was 52.5% at thawing and decreased significantly during the first 3 h of culture in both media (17.1% and 15.4% for SOD and TCM respectively, p < 0:01). SOD supplementation to both media did not influence acrosome integrity during the culture. SOF appeared as more efficient than TCM to preserve stallion sperm viability and motility during in vitro culture.