Authors Schubert, R. ; Strohmeyer, N. ; Bharadwaj, M. ; Ramanathan, S. P. ; Krieg, M. ; Friedrichs, J. ; Franz, C. M. ; Müller, D. J.
Title Assay for characterizing the recovery of vertebrate cells for adhesion measurements by single-cell force spectroscopy
Date 13.10.2014
Number 43491
Abstract Single-cell force spectroscopy (SCFS) is becoming a widely used method to quantify the adhesion of a living cell to a substrate, another cell or tissue. The high sensitivity of SCFS permits determining the contributions of individual cell adhesion molecules (CAMs) to the adhesion force of an entire cell. However, to prepare adherent cells for SCFS, they must first be detached from tissue-culture flasks or plates. EDTA and trypsin are often applied for this purpose. Because cellular properties can be affected by this treatment, cells need to recover before being further characterized by SCFS. Here we introduce atomic force microscopy (AFM)-based SCFS to measure the mechanical and adhesive properties of HeLa cells and mouse embryonic kidney fibroblasts while they are recovering after detachment from tissue-culture. We find that mechanical and adhesive properties of both cell lines recover quickly (<10 min) after detachment using EDTA, while trypsin-detached fibroblasts require >60 min to fully recover. Our assay introduced to characterize the recovery of mammalian cells after detachment can in future be used to estimate the recovery behavior of other adherent cell types.
Publisher FEBS Letters
Citation FEBS Letters 588 (2014) 3639-3648
Tags atomic force microscopy collagen i fibronectin trypsin ethylenediaminetetraacetic acid extracellular matrix

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