Endothelial cell adhesion, signaling, and morphogenesis in fibroblast-derived matrix

Patricia A. Soucy, Lewis Romer

Research output: Contribution to journalArticle

Abstract

Extracellular matrix plays a critical role in cellular development by providing signaling cues that direct morphogenesis. In order to study both the cues that natural matrix provides and endothelial cell responses to that information, human fetal lung fibroblasts were used to produce a fibrous three-dimensional matrix. Following the removal of the fibroblasts by detergent extraction, protein and proteoglycan constituents of the remaining matrix were identified by immunofluorescence and immunoblotting. Matrix components included fibronectin, tenascin-C, collagen I, collagen IV, collagen VI, versican, and decorin. Colocalization analysis suggested that fibronectin was a uniquely distributed matrix protein. Morphology, three-dimensional matrix adhesions, and integrin-mediated signaling during vasculogenesis were then studied in human endothelial cells seeded onto the fibroblast-derived matrix. Elongated morphology and decreased cell area were noted, as compared with cells on fibronectin-coated coverslips. Cell-matrix adhesions contained vinculin, pY397-FAK, and pY410-p130Cas, and all of these colocalized more with fibronectin than tenascin-C, collagen I, or collagen VI. Additionally, the endothelial cells remodeled the fibroblast-derived matrix and formed networks of tubes with demonstrable lumens. Matrix adhesions in these tubes also predominantly colocalized with fibronectin. The pattern of membrane type 1 matrix metalloprotease expression in the endothelial cells suggested its involvement in the matrix remodeling that occurred during tubulogenesis. These results indicated that information in fibroblast-derived matrix promoted vasculogenic behavior.

Original languageEnglish (US)
Pages (from-to)273-283
Number of pages11
JournalMatrix Biology
Volume28
Issue number5
DOIs
StatePublished - Jun 2009

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Morphogenesis
Fibronectins
Cell Adhesion
Collagen
Endothelial Cells
Fibroblasts
Tenascin
Cues
Versicans
Cell-Matrix Junctions
Decorin
Vinculin
Metalloproteases
Proteoglycans
Immunoblotting
Integrins
Detergents
Fluorescent Antibody Technique
Extracellular Matrix
Proteins

Keywords

  • Cell adhesion
  • Endothelial cell
  • Extracellular matrix
  • Fibronectin
  • Phosphotyrosine
  • Tubulogenesis

ASJC Scopus subject areas

  • Molecular Biology

Cite this

Endothelial cell adhesion, signaling, and morphogenesis in fibroblast-derived matrix. / Soucy, Patricia A.; Romer, Lewis.

In: Matrix Biology, Vol. 28, No. 5, 06.2009, p. 273-283.

Research output: Contribution to journalArticle

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