Please use this identifier to cite or link to this item: doi:10.22028/D291-29008
Volltext verfügbar? / Dokumentlieferung
Title: Quantitative analysis of F-actin alterations in adherent human mesenchymal stem cells: Influence of slow-freezing and vitrification-based cryopreservation
Author(s): Müllers, Yannik
Meiser, Ina
Stracke, Frank
Riemann, Iris
Lautenschläger, Franziska
Neubauer, Julia C.
Zimmermann, Heiko
Language: English
Volume: 14
Issue: 1
Publisher/Platform: PLOS
Year of Publication: 2019
Publikation type: Journal Article
Abstract: Cryopreservation is an essential tool to meet the increasing demand for stem cells in medical applications. To ensure maintenance of cell function upon thawing, the preservation of the actin cytoskeleton is crucial, but so far there is little quantitative data on the influence of cryopreservation on cytoskeletal structures. For this reason, our study aims to quantitatively describe cryopreservation induced alterations to F-actin in adherent human mesenchymal stem cells, as a basic model for biomedical applications. Here we have characterised the actin cytoskeleton on single-cell level by calculating the circular standard deviation of filament orientation, F-actin content, and average filament length. Cryo-induced alterations of these parameters in identical cells pre and post cryopreservation provide the basis of our investigation. Differences between the impact of slow-freezing and vitrification are qualitatively analyzed and highlighted. Our analysis is supported by live cryo imaging of the actin cytoskeleton via two photon microscopy. We found similar actin alterations in slow-frozen and vitrified cells including buckling of actin filaments, reduction of F-actin content and filament shortening. These alterations indicate limited functionality of the respective cells. However, there are substantial differences in the frequency and time dependence of F-actin disruptions among the applied cryopreservation strategies; immediately after thawing, cytoskeletal structures show least disruption after slow freezing at a rate of 1°C/min. As post-thaw recovery progresses, the ratio of cells with actin disruptions increases, particularly in slow frozen cells. After 120 min of recovery the proportion of cells with an intact actin cytoskeleton is higher in vitrified than in slow frozen cells. Freezing at 10°C/min is associated with a high ratio of impaired cells throughout the post-thawing culture.
DOI of the first publication: 10.1371/journal.pone.0211382
URL of the first publication:
Link to this record: hdl:20.500.11880/27876
ISSN: 1932-6203
Date of registration: 24-Sep-2019
Faculty: NT - Naturwissenschaftlich- Technische Fakultät
Department: NT - Physik
Professorship: NT - Jun.-Prof. Dr. Franziska Lautenschläger
Collections:SciDok - Der Wissenschaftsserver der Universität des Saarlandes

Files for this record:
There are no files associated with this item.

Items in SciDok are protected by copyright, with all rights reserved, unless otherwise indicated.