Simulation of Cryoprotectant Addition and Removal
By combining the information formulated from the osmotic tolerance limits of boar spermatozoa along with membrane permeability characteristics, optimal procedures for CPA addition and removal can be determined that minimize osmotic volume excursion and maximize cell viability. Because EG permeates the boar spermatozoa plasma membrane more rapidly than does glycerol, there is less volume excursion during its addition and removal. However, because of the high degree of boar sperm sensitivity to osmotic changes, an abrupt single-step addition or removal of EG does exceed the estimated osmotic tolerance limits of boar spermatozoa. Computer simulations indicate that glycerol addition and removal is even more damaging than addition and removal of EG. Buy Asthma Inhalers Online
Therefore, during CPA addition and removal, it will be necessary to add and remove the CPAs over time. Figures 4 and 5 illustrate how boar sperm cell volume excursion could be minimized by extending the time allotted for addition and removal procedures. Volume excursions are maintained well within the boar sperm’s osmotic tolerance limits if glycerol is added drop by drop over a minimum of 5 min, and if EG is added drop by drop over a minimum of 3 min. Removal is similar, with glycerol requiring a 3-min dilution and EG requiring a 1-min dilution.