Assay for characterizing the recovery of vertebrate cells for adhesion measurements by single-cell force spectroscopy

被引:28
作者
Schubert, Rajib [1 ]
Strohmeyer, Nico [1 ]
Bharadwaj, Mitasha [1 ]
Ramanathan, Subramanian P. [1 ]
Krieg, Michael [2 ]
Friedrichs, Jens [3 ]
Franz, Clemens M. [4 ]
Muller, Daniel J. [1 ]
机构
[1] Swiss Fed Inst Technol, Dept Biosyst Sci & Engn, CH-4058 Basel, Switzerland
[2] Stanford Univ, Dept Cellular & Mol Physiol, Stanford, CA 94305 USA
[3] Leibniz Inst Polymer Res Dresden, Inst Biofunct Polymer Mat, D-01069 Dresden, Germany
[4] Karlsruhe Inst Technol, DFG Ctr Funct Nanostruct, D-76131 Karlsruhe, Germany
基金
瑞士国家科学基金会;
关键词
Atomic force microscopy; Collagen I; Fibronectin; Trypsin; Ethylenediaminetetraacetic acid; Extracellular matrix; LIVING CELLS; AFM-CANTILEVERS; INTEGRINS; FIBRONECTIN; CADHERINS; SYSTEMS; RGD; ALPHA-5-BETA-1; RECOGNITION; MICROSCOPE;
D O I
10.1016/j.febslet.2014.06.012
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
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. (C) 2014 Federation of European Biochemical Societies. Published by Elsevier B.V. All rights reserved.
引用
收藏
页码:3639 / 3648
页数:10
相关论文
共 77 条
[1]   Structure, function and pathophysiology of protease activated receptors [J].
Adams, Mark N. ;
Ramachandran, Rithwik ;
Yau, Mei-Kwan ;
Suen, Jacky Y. ;
Fairlie, David P. ;
Hollenberg, Morley D. ;
Hooper, John D. .
PHARMACOLOGY & THERAPEUTICS, 2011, 130 (03) :248-282
[2]  
ALBIGESRIZO C, 1995, J CELL SCI, V108, P3317
[3]   Cell adhesion in embryo morphogenesis [J].
Barone, Vanessa ;
Heisenberg, Carl-Philipp .
CURRENT OPINION IN CELL BIOLOGY, 2012, 24 (01) :148-153
[4]   Deciphering Teneurin Domains That Facilitate Cellular Recognition, Cell-Cell Adhesion, and Neurite Outgrowth Using Atomic Force Microscopy-Based Single-Cell Force Spectroscopy [J].
Beckmann, Jan ;
Schubert, Rajib ;
Chiquet-Ehrismann, Ruth ;
Mueller, Daniel J. .
NANO LETTERS, 2013, 13 (06) :2937-2946
[5]   Discrete interactions in cell adhesion measured by single-molecule force spectroscopy [J].
Benoit, M ;
Gabriel, D ;
Gerisch, G ;
Gaub, HE .
NATURE CELL BIOLOGY, 2000, 2 (06) :313-317
[6]   Measuring cell adhesion forces with the atomic force microscope at the molecular level [J].
Benoit, M ;
Gaub, HE .
CELLS TISSUES ORGANS, 2002, 172 (03) :174-189
[7]   The integrin-actin connection, an eternal love affair [J].
Brakebusch, C ;
Fässler, R .
EMBO JOURNAL, 2003, 22 (10) :2324-2333
[8]   Elongated Membrane Tethers, Individually Anchored by High Affinity α4β1/VCAM-1 Complexes, Are the Quantal Units of Monocyte Arrests [J].
Chu, Calvin ;
Celik, Emrah ;
Rico, Felix ;
Moy, Vincent T. .
PLOS ONE, 2013, 8 (05)
[9]   Calcium signaling [J].
Clapham, David E. .
CELL, 2007, 131 (06) :1047-1058
[10]   Thrombin signalling and protease-activated receptors [J].
Coughlin, SR .
NATURE, 2000, 407 (6801) :258-264