AFM Monitoring the Influence o f Selected Cryoprotectants on Regeneration of Cryopreserved Cells Mechanical Properties

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Authors

GOLAN Martin JELÍNKOVÁ Šárka KRATOCHVÍLOVÁ Irena SKLÁDAL Petr PEŠL Martin ROTREKL Vladimír PŘIBYL Jan

Year of publication 2018
Type Article in Periodical
Magazine / Source Frontiers in Physiology
MU Faculty or unit

Central European Institute of Technology

Citation
Web https://www.frontiersin.org/articles/10.3389/fphys.2018.00804/full
Doi http://dx.doi.org/10.3389/fphys.2018.00804
Keywords cryopreservation; cell stiffness; AFM; fluorescence microscopy; DMSO; PEG
Description 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.
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