Post-thawing survival of motile ram sperm after isolation by layering on protein columns

Post-thawing survival of ram sperm was examined after semen which had been layered on top of isolation columns containing solutions of bovine serum albumin (BSA) in Tris diluent was processed for freezing by the pellet method. Sperm isolated from the bottom of the BSA column had better post-thawing survival than sperm from the top of the column. The efficiency of sperm isolation was affected by the concentration of BSA in the column, the holding time of semen on the column and the concentration of sperm in the layered semen. The best post-thawing survival of sperm occurred when semen diluted to a concentration of 200 x 10(6) sperm/ml was layered on a column of 6% BSA in Tris diluent and the bottom layer of the column was isolated for freezing after two hours' holding time.