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The development of cryoprotectants and cryoprotective solutions
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Murray, Alex (2023) The development of cryoprotectants and cryoprotective solutions. PhD thesis, University of Warwick.
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Official URL: http://webcat.warwick.ac.uk/record=b3945017
Abstract
Cryopreservation is the use of low temperatures to preserve biological materials. This has applications for the transport and storage of medical products such as red blood cells (RBCs) and cell therapies. Cold is damaging to cells because of ice formation, which necessitates the use of cryoprotectants to mitigate this damage. Current cryoprotectants have limited effectiveness, and are mostly based on organic solvents introduced more than 50 years ago, making the development of novel cryoprotectants and cryoprotectant formations a crucial endeavour to enable new therapies. Chapter 1 is an introduction which provides an overview of the principles of cryopreservation. Chapter 2 is a literature review comparing the effectiveness of trehalose to that of other sugars. This informed the choice of sugar to be used in subsequent chapters. It also collects many studies on trehalose in one place, providing a useful resource for cryopreservation research. Chapter 3 details the development of a multi-component cryoprotectant solution for RBCs. This solution is an improvement over current blood preservation methods because it can be rapidly washed out, decreasing time to treatment. Chapter 4 details the design, synthesis, and testing of a trehalose ester as a method to transport trehalose into the intracellular space. Attempts are made to identify conditions under which this molecule can be used as a cryoprotectant. Chapter 5 explores methods for the cryopreservation of Jurkat cells, which are a model for T-cell therapies. First, a multi-component DMSO-free cryoprotectant solution is developed. Then the effect of L-proline on Jurkat cryopreservation is investigated. The Appendix provides preliminary data from a directed evolution experiment where Jurkat cells are exposed to repeated freeze-thaw-grow cycles to select for freeze tolerance. In summary, several approaches to cryopreservation were explored, resulting in the development of a rapid washout cryoprotectant solution for RBCs, a DMSO-free cryoprotectant solution for Jurkat cells, and the generation of data which will be useful for future research.
Item Type: | Thesis (PhD) | ||||
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Subjects: | Q Science > QH Natural history > QH301 Biology Q Science > QP Physiology T Technology > TP Chemical technology |
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Library of Congress Subject Headings (LCSH): | Cryopreservation of organs, tissues, etc, Blood -- Collection and preservation, Erythrocytes, Trehalose, Cryobiology | ||||
Official Date: | January 2023 | ||||
Dates: |
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Institution: | University of Warwick | ||||
Theses Department: | School of Life Sciences | ||||
Thesis Type: | PhD | ||||
Publication Status: | Unpublished | ||||
Supervisor(s)/Advisor: | Gibson, Matthew I. | ||||
Format of File: | |||||
Extent: | 277 pages : illustrations | ||||
Language: | eng |
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