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Post-Thaw Culture and Measurement of Total Cell Recovery Is Crucial in the Evaluation of New Macromolecular Cryoprotectants
journal contribution
posted on 2020-06-22, 14:34 authored by Kathryn
A. Murray, Matthew I. GibsonThe
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.
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biomaterials researchstudy post-thawTotal Cell RecoveryNew Macromolecular CryoprotectantsviabilitypositiveSeveral systemspost-thaw outcomescell biologycell-based therapiescell recoverynon-cryoprotective polymerspost-thaw culturing timepost-thaw time scalesPost-Thaw Cultureice-binding proteinscryoprotectantPost-thaw culture timeDMSOcryoprotective agents
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