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Post-thaw culture and measurement of total cell recovery is crucial in the evaluation of new macromolecular cryoprotectants
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Murray, Kathryn and Gibson, Matthew I. (2020) Post-thaw culture and measurement of total cell recovery is crucial in the evaluation of new macromolecular cryoprotectants. Biomacromolecules, 21 (7). pp. 2864-2873. doi:10.1021/acs.biomac.0c00591 ISSN 1525-7797.
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WRAP-Post-thaw-culture-measurement-total-cell-recovery-Gibson-2020.pdf - Accepted Version - Requires a PDF viewer. Download (4Mb) | Preview |
Official URL: https://doi.org/10.1021/acs.biomac.0c00591
Abstract
The storage and transport of cells is a fundamental technology which underpins cell biology, biomaterials research, and emerging cell-based therapies. Inspired by antifreeze and ice-binding proteins in extremophiles, macromolecular (polymer) cryoprotectants are emerging as exciting biomaterials to enable the reduction and/or replacement of conventional cryoprotective agents such as DMSO. Here, we critically study post-thaw cellular outcomes upon addition of macromolecular cryoprotectants to provide unambiguous evidence that post-thaw culturing time and a mixture of assays are essential to claim a positive outcome. In particular, we observe that only measuring the viability of recovered cells gives false positives, even with non-cryoprotective polymers. Several systems gave apparently high viability but very low total cell recovery, which could be reported as a success but in practical applications would not be useful. Post-thaw culture time is also shown to be crucial to enable apoptosis to set in. Using this approach we demonstrate that polyampholytes (a rapidly emerging class of cryoprotectants) improve post-thaw outcomes across both measures, compared to poly(ethylene glycol), which can give false positives when only viability and short post-thaw time scales are considered. This work will help guide the discovery of new macromolecular cryoprotectants and ensure materials which only give positive results under limited outcomes can be quickly identified and removed.
| Item Type: | Journal Article | ||||||||||||||||||
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| Subjects: | Q Science > QH Natural history Q Science > QP Physiology |
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| Divisions: | Faculty of Science, Engineering and Medicine > Science > Chemistry | ||||||||||||||||||
| Library of Congress Subject Headings (LCSH): | Macromolecules -- Research, Cytology -- Research, Cryobiology | ||||||||||||||||||
| Journal or Publication Title: | Biomacromolecules | ||||||||||||||||||
| Publisher: | American Chemical Society | ||||||||||||||||||
| ISSN: | 1525-7797 | ||||||||||||||||||
| Official Date: | 13 July 2020 | ||||||||||||||||||
| Dates: |
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| Volume: | 21 | ||||||||||||||||||
| Number: | 7 | ||||||||||||||||||
| Page Range: | pp. 2864-2873 | ||||||||||||||||||
| DOI: | 10.1021/acs.biomac.0c00591 | ||||||||||||||||||
| Status: | Peer Reviewed | ||||||||||||||||||
| Publication Status: | Published | ||||||||||||||||||
| Reuse Statement (publisher, data, author rights): | “This document is the Accepted Manuscript version of a Published Work that appeared in final form in Biomacromolecules, copyright © American Chemical Society after peer review and technical editing by the publisher. To access the final edited and published work see [insert ACS Articles on Request author-directed link to Published Work, see http://pubs.acs.org/page/policy/articlesonrequest/index.html].” | ||||||||||||||||||
| Access rights to Published version: | Restricted or Subscription Access | ||||||||||||||||||
| Date of first compliant deposit: | 3 July 2020 | ||||||||||||||||||
| Date of first compliant Open Access: | 22 June 2021 | ||||||||||||||||||
| RIOXX Funder/Project Grant: |
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