Wiggin H B, Almquist J O
J Dairy Sci. 1975 Mar;58(3):416-9. doi: 10.3168/jds.S0022-0302(75)84580-2.
Twelve ejaculates were used in a central composite experiment to test 15 combinations of glycerol (7, 9, 11, 13, or 15%), glycerol equilibration times (1, 2, 4, 8, or 16 h) and thawing rates (water at 35 C for 15 s, 50 C for 13 s, 65 C for 11 s, 80 C for 9 s, or 95 C for 7 s). Semen was diluted in heated skim milk-glycerol, packaged in .3-ml. Continental U.S. straws and frozen in liquid nitrogen vapor. Based on post-thaw progressive sperm motility after storage at -196 C for 9 to 11 days, estimated optima from multiple regression were 10.7% for glycerol, 2.0 h for glycerol equilibration time, and 76 C for thawing bath temperature. Only the linear effect for each variable was significant. Much faster thawing rates and shorter glycerol equilibration times than those for freezing bull spermatozoa in glass ampules should be used for maximum post-thaw sperm motility in straws.
在一项中心复合实验中，使用了12份射精样本，以测试甘油浓度（7%、9%、11%、13%或15%）、甘油平衡时间（1小时、2小时、4小时、8小时或16小时）和解冻速率（35℃水浴15秒、50℃水浴13秒、65℃水浴11秒、80℃水浴9秒或95℃水浴7秒）的15种组合。精液用加热的脱脂乳 - 甘油稀释，装入0.3毫升的美国大陆规格细管中，并在液氮蒸汽中冷冻。根据在 -196℃储存9至11天后解冻后的精子渐进性运动能力，多元回归估计的最佳值为甘油浓度10.7%、甘油平衡时间2.0小时、解冻浴温度76℃。每个变量仅线性效应显著。为使细管中精子解冻后运动能力达到最大值，应采用比在玻璃安瓿中冷冻公牛精子更快的解冻速率和更短的甘油平衡时间。
