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Cellular osmotic properties and cellular responses to cooling Open Access


Other title
interrupted freezing
cryobiological simulations
osmotic properties
Type of item
Degree grantor
University of Alberta
Author or creator
Ross-Rodriguez, Lisa Ula
Supervisor and department
McGann, Locksley E. (Laboratory Medicine and Pathology)
Elliott, Janet A.W. (Chemical and Materials Engineering)
Examining committee member and department
Hugh, Judith (Laboratory Medicine and Pathology)
Churchill, Thomas (Surgery)
Critser, John (Veterinary Pathobiology)
Korbutt, Gregory (Surgery)
Medical Sciences- Laboratory Medicine and Pathology

Date accepted
Graduation date
Doctor of Philosophy
Degree level
Recent advances in the fundamental theories in cryobiology using thermodynamic principles have created new opportunities for innovative methodologies in cryobiology. This thesis tested the hypothesis that calculated indicators of the two-factor hypothesis of cryoinjury, depending on cellular osmotic properties, will describe outcomes of cryobiological experiments. In addition, this thesis demonstrated that knowledge gained from improved descriptions of cellular osmotic parameters allows better understanding of cryoinjury and cryoprotection. The main objective of this thesis was to develop approaches using simulations that can be applied to development of cryopreservation procedures for cell types of interest for therapies. In order for this approach to be successful, a method to more accurately describe the osmotic solution properties of the cell (i.e. osmolality as a function of molality for the cytoplasm) was developed. Also, in-depth examination into the correlation between predictions of the two types of cryoinjury and measured post-thaw biological outcomes was required. The work presented in this thesis has shown that simulations, based on cell-specific osmotic characteristics, and coupled with interrupted cooling procedures can be used to determine conditions that minimize the two identified damaging factors in cryopreservation. Based on results from this research, both intracellular supercooling and osmolality, as indicators of intracellular ice formation and solution effects injury, respectively, should be calculated when attempting to compare simulations with biological experimentation. This thesis has also shown a novel method of obtaining the solution properties (i.e. osmolality as a function of molality) of the cytoplasm of living cells using equilibrium cell volume measurements. Using these newly calculated parameters, this research also demonstrated the magnitude of error introduced by making dilute solution assumptions of the solution properties in cellular responses to low temperatures, including simulations of interrupted freezing procedures. Overall, the research work presented in this thesis has extended the approach to cryopreservation to include the properties of the cell and the physical conditions of the freezing environment, which was only possible through the linkage between biological experimentation and simulations.
Permission is hereby granted to the University of Alberta Libraries to reproduce single copies of this thesis and to lend or sell such copies for private, scholarly or scientific research purposes only. Where the thesis is converted to, or otherwise made available in digital form, the University of Alberta will advise potential users of the thesis of these terms. The author reserves all other publication and other rights in association with the copyright in the thesis and, except as herein before provided, neither the thesis nor any substantial portion thereof may be printed or otherwise reproduced in any material form whatsoever without the author's prior written permission.
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