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 - 1997

Fingerprint

Integrins
Smooth Muscle Myocytes
Cell Movement
Vitronectin
Muscle
Assays
Osteopontin
Cells
Chemotaxis
Monoclonal antibodies
Extracellular Matrix Proteins
Chemotactic Factors
Cell adhesion
Endothelial cells
Antibodies
Screening
Monoclonal Antibodies
Proteins
Pathologic Processes
Molecules

Keywords

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

ASJC Scopus subject areas

  • Cell Biology

Cite this

An improved method for the quantitation of cellular migration : Role of αvβ3 integrin in endothelial and smooth muscle cell migration. / Bozarth, Jeffrey M.; Penno, Margaret B.; Mousa, Shaker A.

In: Methods in Cell Science, Vol. 19, No. 3, 1997, p. 179-187.

Research output: Contribution to journalArticle

@article{21a55e3f8f2a462dab7bc094cc0a8c17,
title = "An improved method for the quantitation of cellular migration: Role of αvβ3 integrin in endothelial and smooth muscle cell migration",
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.",
keywords = "αvβ3 integrin, Chemotaxis, Endothelial cell migration, Fluorescence measurement, Quantitation, Smooth muscle migration",
author = "Bozarth, {Jeffrey M.} and Penno, {Margaret B.} and Mousa, {Shaker A.}",
year = "1997",
doi = "10.1023/A:1009789416463",
language = "English (US)",
volume = "19",
pages = "179--187",
journal = "Cytotechnology",
issn = "0920-9069",
publisher = "Springer Netherlands",
number = "3",

}

TY - JOUR

T1 - An improved method for the quantitation of cellular migration

T2 - Role of αvβ3 integrin in endothelial and smooth muscle cell migration

AU - Bozarth, Jeffrey M.

AU - Penno, Margaret B.

AU - Mousa, Shaker A.

PY - 1997

Y1 - 1997

N2 - 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.

AB - 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.

KW - αvβ3 integrin

KW - Chemotaxis

KW - Endothelial cell migration

KW - Fluorescence measurement

KW - Quantitation

KW - Smooth muscle migration

UR - http://www.scopus.com/inward/record.url?scp=0031417110&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0031417110&partnerID=8YFLogxK

U2 - 10.1023/A:1009789416463

DO - 10.1023/A:1009789416463

M3 - Article

AN - SCOPUS:0031417110

VL - 19

SP - 179

EP - 187

JO - Cytotechnology

JF - Cytotechnology

SN - 0920-9069

IS - 3

ER -