This article is published in Journal of Molecular Recognition as part of the virtual Special Issue ‘AFM BioMed Shanghai 2013, edited by Jun Hu, SINAP, China and Pierre Parot and Jean-Luc Pellequer, CEA, France’.
Special Issue Article
Investigating differential cell-matrix adhesion by directly comparative single-cell force spectroscopy†
Article first published online: 1 OCT 2013
Copyright © 2013 John Wiley & Sons, Ltd.
Journal of Molecular Recognition
Volume 26, Issue 11, pages 578–589, November 2013
How to Cite
Dao, L., Gonnermann, C. and Franz, C. M. (2013), Investigating differential cell-matrix adhesion by directly comparative single-cell force spectroscopy. J. Mol. Recognit., 26: 578–589. doi: 10.1002/jmr.2303
- Issue published online: 1 OCT 2013
- Article first published online: 1 OCT 2013
- Manuscript Accepted: 31 JUL 2013
- Manuscript Revised: 24 JUL 2013
- Manuscript Received: 29 JUN 2013
- single-cell force spectroscopy;
- cell adhesion;
- microcontact printing;
- differential adhesion;
Tissue-embedded cells are often exposed to a complex mixture of extracellular matrix (ECM) molecules, to which they bind with different cell adhesion receptors and affinities. Differential cell adhesion to ECM components is believed to regulate many aspects of tissue function, such as the sorting of specific cell types into different tissue compartments or ECM niches. In turn, aberrant switches in cell adhesion preferences may contribute to cell misplacement, tissue invasion, and metastasis. Methods to determine differential adhesion profiles of single cells are therefore desirable, but established bulk assays usually only test cell population adhesion to a single type of ECM molecule. We have recently demonstrated that atomic force microscopy-based single-cell force spectroscopy (SCFS), performed on bifunctional, microstructured adhesion substrates, provides a useful tool for accurately quantitating differential matrix adhesion of single Chinese hamster ovary cells to laminin and collagen I. Here, we have extended this approach to include additional ECM substrates, such as bifunctional collagen I/collagen IV surfaces, as well as adhesion-passivated control surfaces. We investigate differential single cell adhesion to these substrates and analyze in detail suitable experimental conditions for comparative SCFS, including optimal cell-substrate contact times and the impact of force cycle repetitions on single cell adhesion force statistics. Insight gained through these experiments may help in adapting this technique to other ECM molecules and cell systems, making directly comparative SCFS a versatile tool for comparing receptor-mediated cell adhesion to different matrix molecules in a wide range of biological contexts. Copyright © 2013 John Wiley & Sons, Ltd.