Bailey, Trisha L., Stubbs, Christopher, Murray, Kathryn, Tomás, Ruben M. F., Otten, Lucienne and Gibson, Matthew I. (2019) Data for Synthetically scalable poly(ampholyte) which dramatically enhances cellular cryopreservation. [Dataset]

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Abstract

The storage and transport of frozen cells underpin the emerging/existing cell-based therapies and are used in every biomedical research lab globally. The current gold-standard cryoprotectant dimethyl sulfoxide (DMSO) does not give quantitative cell recovery in suspension or in two-dimensional (2D) or three-dimensional (3D) cell models, and the solvent and cell debris must be removed prior to application/transfusion. There is a real need to improve this 50-year-old method to underpin emerging regenerative and cell-based therapies. Here, we introduce a potent and synthetically scalable polymeric cryopreservation enhancer which is easily obtained in a single step from a low cost and biocompatible precursor, poly(methyl vinyl ether-alt-maleic anhydride). This poly(ampholyte) enables post-thaw recoveries of up to 88% for a 2D cell monolayer model compared to just 24% using conventional DMSO cryopreservation. The poly(ampholyte) also enables reduction of [DMSO] from 10 wt % to just 2.5 wt % in suspension cryopreservation, which can reduce the negative side effects and speed up post-thaw processing. After thawing, the cells have reduced membrane damage and faster growth rates compared to those without the polymer. The polymer appears to function by a unique extracellular mechanism by stabilization of the cell membrane, rather than by modulation of ice formation and growth. This new macromolecular cryoprotectant will find applications across basic and translational biomedical science and may improve the cold chain for cell-based therapies.

Item Type:	Dataset
Subjects:	Q Science > QD Chemistry Q Science > QH Natural history Q Science > QP Physiology
Divisions:	Faculty of Science, Engineering and Medicine > Science > Chemistry
Type of Data:	Experimental data
Library of Congress Subject Headings (LCSH):	Cryopreservation of organs, tissues, etc., Cells -- Cryopreservation, Frozen erythrocytes, Polyampholytes
Journal or Publication Title:	Biomacromolecules
Publisher:	Department of Chemistry
ISSN:	1525-7797
Official Date:	31 July 2019
Dates:	Date Event 31 July 2019 Published
DOI:	10.1021/acs.biomac.9b00681
Status:	Not Peer Reviewed
Publication Status:	Published
Media of Output (format):	.xlsx
Access rights to Published version:	Open Access
Copyright Holders:	University of Warwick
Description:	Differential scanning calorimetry includes data used to calculate ice crystallization enthalpy and melting temperature, along with full DSC traces and is located in the “DSC” folder. Ice recrystallization data was used to calculate the mean grain size is located in the “IRI” folder. Cytotoxicity data was used to calculate alamar blue reduction for P2 and DMSO and each file is located in the “mammalian cell cytotoxicity” folder. Monolayer and suspension recoveries for cryopreserved A549 is located in the “mammalian cell freezing” folder, subfolder “A549 cell freezing” and includes P2 concentration recoveries, lower DMSO concentrations with P2, and molecular weight effects of polyampholyte. Monolayer recoveries for cryopreserved MC-3T3 and Neuro-2a cells is located in the “mammalian cell freezing” folder, subfolder “Neuro-2a & MC-3T3 cell freezing”. Fold change data for all three cell lines can be found in the “mammalian cell freezing” folder. Post-freeze A549 growth rate data is located in the “mammalian cell growth rates” folder. Membrane kinetics for incubated A549 cells is located in the “mammalian cell membrane kinetics” folder. Live/dead graphical representation from post-freeze staining data is located in the “mammalian cell post-freeze imaging” folder. Polyampholyte NMR and FITR data is located in the “polymer characterisation” folder. Red blood cell incubation results, osmotic fragility, polyampholyte freezing, and polymer comparisons data is located in the “red blood cell freezing” folder.
RIOXX Funder/Project Grant:	Project/Grant ID RIOXX Funder Name Funder ID 638661 FP7 Ideas: European Research Council http://dx.doi.org/10.13039/100011199 789182 FP7 Ideas: European Research Council http://dx.doi.org/10.13039/100011199 EP/L015307/1RT [EPSRC] Engineering and Physical Sciences Research Council http://dx.doi.org/10.13039/501100000266 105627/Z/14/Z Wellcome Trust http://dx.doi.org/10.13039/100010269 ALERT14 award BB/M01228X/1 [BBSRC] Biotechnology and Biological Sciences Research Council http://dx.doi.org/10.13039/501100000268 105627/Z/14/Z University of Warwick http://dx.doi.org/10.13039/501100000741
Related URLs:	Related item in WRAP Other
Contributors:	Contribution Name Contributor ID Depositor Gibson, Matthew I. 32015

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