These data were later applied to resolve previous apparent discrepancies between theoretical predictions and empirical outcomes. They were then used to develop more optimal cryopreservation protocols for human spermatozoa . The same approach to fundamental cryobiology should be used as a paradigm for the optimization of boar spermatozoa cryopreservation.
Theoretical computer simulations of intracellular water volume flux during cooling and warming indicate that boar spermatozoa respond similarly in the presence of glycerol and EG at relatively low cooling rates. The data suggest that boar spermatozoa should tolerate cooling rates of 10°C/ min and 100°C/min in combination with warming rates of 1200°C/min with minimal damage. However, excessively higher cooling rates (e.g., 1000°C/min) in the presence of EG do result in potentially damaging cell volume excursions. Similar experimental findings of damaging cell volume excursions upon cooling and warming have been previously reported in other cell types such Chinese hamster ovary cells . In addition, the theoretical simulations indicate that cell loss during cooling and warming is not primarily a result of intracellular ice formation. The probability of intracellular ice formation in boar sperm is less than 5%, in the presence of either glycerol or EG, when using cooling rates of 10°C/min, 100°C/min, and 1000°C/min.