Publication details

Cryopreserved Cells Regeneration Monitored by Atomic Force Microscopy and Correlated With State of Cytoskeleton and Nuclear Membrane

Authors

GOLAN Martin PŘIBYL Jan PEŠL Martin JELÍNKOVÁ Šárka AĆIMOVIĆ Ivana JAROŠ Josef ROTREKL Vladimír FALK Martin SEFC Ludek SKLÁDAL Petr KRATOCHVILOVA Irena

Year of publication 2018
Type Article in Periodical
Magazine / Source IEEE TRANSACTIONS ON NANOBIOSCIENCE
MU Faculty or unit

Central European Institute of Technology

Citation
Doi http://dx.doi.org/10.1109/TNB.2018.2873425
Keywords Atomic forcemicroscopy; cell surface stiffness; cryopreservation; cryopreservedcells reconstruction
Description Atomic force microscopy (AFM) helps to describe and explain the mechanobiological properties of living cells on the nanoscale level under physiological conditions. The stiffness of cells is an important parameter reflecting cell physiology. Here, we have provided the first study of the stiffness of cryopreserved cells during post- thawing regeneration using AFM combined with confocal fluorescence microscopy. We demonstrated that the nonfrozen cell stiffness decreased proportionally to the cryoprotectant concentration in the medium. AFM allowed us to map cell surface reconstitution in real time after a freeze/thaw cycle and to monitor the regeneration processes at different depths of the cell and even different parts of the cell surface (nucleus and edge). Fluorescence microscopy showed that the cytoskeleton in fibroblasts, though damaged by the freeze/thaw cycle, is reconstructed after long-termplating. Confocalmicroscopy confirmed that structural changes affect the nuclear envelopes in cryopreserved cells. AFM nanoindentation analysis could be used as a noninvasive method to identify cells that have regenerated their surface mechanical properties with the proper dynamics and to a sufficient degree. This identification can be important particularly in the field of in vitro fertilization and in future cell-based regeneration strategies.
Related projects: