Quantitative Measurement of Cell Adhesion Using Centrifugal Force

David R. McClay1, Philip L. Hertzler1

1 Duke University, Durham, North Carolina
Publication Name:  Current Protocols in Cell Biology
Unit Number:  Unit 9.2
DOI:  10.1002/0471143030.cb0902s00
Online Posting Date:  May, 2001
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Abstract

In order to study the biophysical forces involved in cell‐substrate (or cell‐cell) adhesion, it is necessary to measure the strength of adhesion. Two questions can be addressed using the centrifugal cell adhesion assay provided in this unit: what is the ligand‐receptor affinity for cells adhering at 4oC and what is the strength of the ATP‐dependent processes that strengthen adhesion at 37oC. In both cases, the strength of adhesion is measured as the resistance to disruption in the presence of a measured centrifugal force.

     
 
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Table of Contents

  • Basic Protocol 1: Centrifugation Cell Adhesion Assay
  • Commentary
  • Literature Cited
  • Figures
     
 
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Materials

Basic Protocol 1: Centrifugation Cell Adhesion Assay

  Materials
  • Substrate molecule of interest in PBS
  • 40 mg/ml BSA (fraction V; Sigma) in PBS
  • PBS ( appendix 2A)
  • Tissue culture medium without FBS
  • Dissociated cells of interest, suspended at 5–10 × 105/ml in calcium‐free physiological solution (see , discussion of suitable media)
  • Clear packing tape (3M Scotch 375, 4.8 cm wide, although any clear packing tape will probably work)
  • Microtiter plate support template
  • Low‐speed refrigerated cell centrifuge with microtiter plate carrier
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Figures

Videos

Literature Cited

Literature Cited
   Burdsal, C., Alliegro, M.C., and McClay, D.R. 1991. Echinonectin as a substrate for adhesion during development of the sea urchin embryo. Dev. Biol. 144:327‐334.
   Burdsal, C.A., Lotz, M.M., Miller, J., and McClay, D.R. 1994. A quantitative switch in integrin expression accompanies differentiation of F9 cells treated with retinoic acid. Dev. Dynamics 201:344‐353.
   Dembo, M. and Bell, G.I. 1987. The thermodynamics of cell adhesion. Curr. Top. Membr. Transp. 29:71‐89.
   Lotz, M.M., Burdsal, C.A., Erickson, H.P., and McClay, D.R. 1989. Cell adhesion to fibronectin and tenascin: Quantitative measurements of initial binding and subsequent strengthening response. J. Cell Biol. 109:1795‐1805.
   McClay, D.R., Wessel, G.M., and Marchase, R.B. 1981. Intercellular recognition: Quantitation of initial binding events. Proc. Natl. Acad. Sci. U.S.A. 78:4975‐4979.
   Rich, A.M. 1978. Substratum amd solution parameters of cell adhesion. Ph.D. thesis. University of North Carolina, Chapel Hill.
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