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.
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