An improved method for the quantitation of cellular migration: Role of αvβ3 integrin in endothelial and smooth muscle cell migration

Jeffrey M. Bozarth, Margaret B. Penno, Shaker A. Mousa

Research output: Contribution to journalArticle

Abstract

The present study was undertaken to develop a simple and improved method for the accurate quantitation of cellular migration and to examine the role of αvβ3 integrins in different cellular migration. Using our newly developed micro-volume chemotaxis assay, we developed an improved quantitative method to measure in vitro chemotaxis of smooth muscle or endothelial cells toward different extracellular matrix proteins. The convenience in setup and counting of migrated cells using this method allows for large capacity screening and for various research applications with other cells as well. The signal. to noise ratios were in the range of 10/1, along with about 10-20% intra- or inter-assay variabilities. Using this method, we have determined that either vitronectin at 0.4 μg/well or osteopontin at 0.4 μg/well are selective αvβ3 chemoattractants for endothelial or smooth muscle cells (0.5 x 105 cells/well). Additionally, a selective αvβ3 small molecule peptiddomimetic, monoclonal antibody LM609, or an anti-β3 (αvβ3/αIIβ3) antibody, c7E3 demonstrated maximal inhibition of cellular migration toward vitronectin or osteopontin. These data suggest the potential utility of this method in assessing the role of various mechanisms in cellular migration and also suggests the potential implication of an αvβ3 antagonist in blocking pathological processes involving endothelial or smooth muscle cell adhesion/migration.

Original languageEnglish (US)
Pages (from-to)179-187
Number of pages9
JournalMethods in Cell Science
Volume19
Issue number3
DOIs
StatePublished - Dec 1 1997

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Keywords

  • Chemotaxis
  • Endothelial cell migration
  • Fluorescence measurement
  • Quantitation
  • Smooth muscle migration
  • αvβ3 integrin

ASJC Scopus subject areas

  • Cell Biology

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