Theoretical analysis of efficiency of rapid two-step freezing regimens
Keywords:
non-linear cooling regimens, cryodamage, intracellular solution saturation, intracellular crystallization, Saccharomyces cerevisiaeAbstract
On the basis of traditional concepts on phase transition kinetics during cooling of multi-component solutions
and transmembrane transfer of substances we developed the quantitative model of cryodamage of yeast-like
fungi Saccharomyces cerevisiae suspension in the cryoprotective solution in non-linear cooling regimens at its crystallization stage. The designed theoretical model is verified by the experiments, testifying to the adequacy of the proposed model concept about cryodamage of cells at the stage of cell suspension crystallization, in a particular solution of “dimethyl sulfoxide (10% v/v) – sodium chloride (0.135 M) – water” under exponential cooling regimen. The mechanism of the cryoprotective effect of rapid two-step cooling was shown to have no substantial differences from optimal from the point of view of the two-factor theory of cryodamage coolingegimen with a constant cooling rate. It is established that the optimal non-linear cooling regimen of yeast-like fungi cells Saccharomyces cerevisiae could be implemented by plunging of the container carrying the cell suspension into the liquid-cooled down to -35ºC for 20-25 minutes with following rapid cooling in liquid nitrogen.
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