Cryopreservation of human sperm using rapid cooling rates

The effect of cryopreservation on human spermatozoa has been investigated during the past few decades. The majority of current cryopreservation protocols are carried-out using low cooling rates. However, theoretical calculations have shown that for human spermatozoa an optimal cooling rate is about 7,000 C/min. In our work we have studied the effect of cryopreservation with high cooling rates, variation of osmolarity of the cryoprotectant medium and the glycerol content. The results of experiments have demonstrated that within the range of high cooling rates, after thawing the dependency of sperm survival on the cooling rate has a maximum recovery at 2,500-3,300 C/min in moderately hyperosmolar medium, containing 4-5% glycerol. When decreasing the cooling rate down to 1,750-2,500 C/min there was a statistically significant reduction in sperm motility. Using the cooling rate of 8,000-11,000 C /min only a small percentage of spermatozoa retained their motility.