Cellular Response of Mouse Oocytes to Freezing Stress: Prediction of Intracellular Ice Formation

Abstract

Successful protocols for cryopreservation of living cells can be designed if the physicochemical conditions to preclude intracellular ice formation (IIF) can be defined. Unfortunately, all attempts to predict the probability of IIF have met with very limited success. In this study, an analytical model is developed to predict ice formation inside mouse oocytes subjected to a freezing stress. According to the model, IIF is catalyzed heterogeneously by the plasma membrane (i.e., surface catalyzed nucleation, SCN). A local site on the plasma membrane is assumed to become an ice nucleator in the presence of the extracellular ice via its effects on the membrane. This interaction is characterized by the contact angle between the plasma membrane and the ice cluster. In addition, IIF is assumed to be catalyzed at temperatures below -30° C by intracellular particles distributed throughout the cell volume (i.e., volume catalyzed nucleation, VCN). In the present study, these two distinctly coupled modes of IIF, especially SCN, are applied to various experimental protocols from mouse oocytes. Excellent agreement between predictions and observations suggests that the proposed model of IIF is adequate.