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Kinetics of coupling water and cryoprotectant transport across cell membranes and applications to cryopreservation.

Thermodynamic and kinetic models can provide a wealth of information on the physical response of living cells and tissues experiencing cryopreservation procedures. Both isothermal and nonisothermal models have been proposed so far, accompanied by experimental verification and cryoapplications. But the cryoprotective solution is usually assumed to be dilute and ideal in the models proposed in the literature. Additionally, few nonisothermal models are able to couple the transmembrane transport of water and cryoprotectant during cooling and warming of cells. To overcome these limitations, this study develops a whole new set of equations that can quantify the cotransport of water and cryoprotectant across cell membranes in the nondilute and nonideal solution during the freezing and thawing protocols. The new models proposed here can be simplified into ones consistent with the classic models if some specific assumptions are included. For cryobiological practice, they are applied to predict the volumetric change for imprinting control region (ICR) mouse spermatozoa and human corneal keratocytes in the freezing protocol. The new models can determine the intracellular concentration of cryoprotectant more precisely than others by abandoning the assumptions such as dilute and ideal solutions and nonpermeability of membranes to cryoprotectant. Further, the findings in this study will offer new insights into the physical response of cells undergoing cryopreservation.