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Permanent link (DOI): https://doi.org/10.7939/R38H9X

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Membrane and Mitochondrial Responses to Cryobiological Conditions Open Access

Descriptions

Other title
Subject/Keyword
Flow Cytometry
Cryobiology
Mitochondria
Plasma Membrane
Type of item
Thesis
Degree grantor
University of Alberta
Author or creator
Reardon, Anthony J
Supervisor and department
McGann, Locksley (Laboratory Medicine and Pathology)
Examining committee member and department
Elliott, Janet (Chemical and Materials Engineering)
Churchill, Thomas (Surgery)
Acker, Jason (Laboratory Medicine and Pathology)
Department
Medical Sciences- Laboratory Medicine and Pathology
Specialization

Date accepted
2013-01-04T15:37:48Z
Graduation date
2013-06
Degree
Master of Science
Degree level
Master's
Abstract
The success of cellular cryopreservation is indicated by the state of post-thawed cells. Using human umbilical vein endothelial cells as a model, the cryobiological response to interrupted cooling protocols was evaluated with a combination of fluorescence microscopy and flow cytometry. Cryoinjured cells were identified using fluorescence intensity with flow cytometry, where light scatter gating strategies were shown to have limitations. Plasma membrane integrity has become the standard assay for post-thaw cell survival. This investigation provided a direct comparison of the integrity of the plasma membrane with the functional state of mitochondria in frozen-thawed cells. A disconnect was found between these two sites of cryoinjury: at specific low temperature conditions cells with depolarized mitochondria still had an intact plasma membrane. This result indicates that different mechanisms may affect each of these sites of cryoinjury. These findings are of interest in understanding the mechanisms of cryoinjury required to minimize cell damage during cryopreservation.
Language
English
DOI
doi:10.7939/R38H9X
Rights
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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