AFM Monitoring the Influence of Selected Cryoprotectants on Regeneration of Cryopreserved Cells Mechanical Properties.
Front. Physiol.. 2018-06-29; 9:
DOI: 10.3389/fphys.2018.00804
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Golan M(1), Jelinkova S(2)(3), Kratochvílová I(1), Skládal P(4), Pešl M(2)(3)(5), Rotrekl V(2)(3), Pribyl J(4).
Author information:
(1)Department of Analysis of Functional Materials, Institute of Physics, Academy of Sciences Czech Republic, Prague, Czechia.
(2)Department of Biology, Faculty of Medicine, Masaryk University, Brno, Czechia.
(3)International Clinical Research Center, St. Anne’s University Hospital, Brno, Czechia.
(4)Central European Institute of Technology, Masaryk University, Brno, Czechia.
(5)First Department of Internal Medicine/Cardioangiology, Masaryk University, Brno, Czechia.
Cryopreservation of cells (mouse embryonic fibroblasts) is a fundamental task for
wide range of applications. In practice, cells are protected against damage
during freezing by applications of specific cryoprotectants and freezing/melting
protocols. In this study by using AFM and fluorescence microscopy we showed how
selected cryoprotectants (dimethyl sulfoxide and polyethylene glycol) affected
the cryopreserved cells mechanical properties (stiffness) and how these
parameters are correlated with cytoskeleton damage and reconstruction. We showed
how cryopreserved (frozen and thawed) cells’ stiffness change according to type
of applied cryoprotectant and its functionality in extracellular or intracellular
space. We showed that AFM can be used as technique for investigation of
cryopreserved cells surfaces state and development ex vivo. Our results offer a
new perspective on the monitoring and characterization of frozen cells recovery
by measuring changes in elastic properties by nanoindentation technique. This may
lead to a new and detailed way of investigating the post-thaw development of
cryopreserved cells which allows to distinguish between different cell parts.