A novel model capable of quantitatively describing and predicting intracellular ice formation (IIF) as a function of temperature in a cell population characterized by a size distribution is proposed. The model overcomes the classical approach which takes into account a population of identically sized cells. The size distribution dynamics of a cell population in response to water osmosis and IIF occurrence during the cooling stage of a standard cryopreservation protocol without using cryoprotective agent (CPA) is simulated by means of a suitable population balance approach. Specifically, the model couples the classical water transport equation developed by Mazur1 to the quantitative description of nucleation and diffusion-limited growth of ice crystals in the framework of a 1-D population balance equation (PBE). It is found that IIF temperature depends on the cell size, i.e., it is higher for larger cells. Correspondingly, the probability of IIF (PIIF) results to be dependent on the initial size distribution of the cell population. Model's parameters related to the osmotic behavior of the cell population and to IIF kinetics are obtained by comparison between theoretical results and suitable experimental data of isolated rat hepatocytes available in the literature. Model reliability is successfully verified by predicting the experimental data of PIIF at different, constant cooling rates with better accuracy as compared to the theoretical approaches available in the literature. © 2009 American Institute of Chemical Engineers AIChE J, 2010
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