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Cryopreservation of Human Sperm: Effect of Cooling Rate on Intracellular Ice Formation

The effect of cooling rate on the intracellular ice formation for human spermatozoa have been studied and a correlation formula is established in this study. It is found that after a certain cooling rate, termed as optimal cooling rate in this study, the volume of intracellular ice increases exponentially with increase in the cooling rate. The cellular response of human sperm cells to different coolin g rates, varying between 5°C/min and 300°C/min, is studied using the diffusion limited transport model. The membrane permeability parameters are taken from the published literature. For a cooling rate lower than the optimal value, almost all intracellular water flows out to t he extracellular compartment through the cell membrane leaving behind only the osmotically inactive cell volume inside the cell. For the calculation of the amount of intracellular ice formation, it has been assumed that the amount of water trapped inside the human sperm at -19°C will finally form intracellular ice with sufficient super-cooling.