Endothelial cell differentiation into capillary-like structures in response to tumour cell conditioned medium: a modified chemotaxis chamber assay

Br J Cancer. 1995 Apr;71(4):770-5. doi: 10.1038/bjc.1995.149.

Abstract

We have developed a modified chemotaxis chamber assay in which bovine aortic endothelial (BAE) cells degrade Matrigel basement membrane and migrate and form capillary-like structures on type I collagen. This capillary formation occurs in the presence of conditioned media from highly metastatic tumour cell lines, such as B16F10 murine melanoma or MDA-MD-231 human breast adenocarcinoma, but not in the presence of conditioned medium (CM) from the less invasive B16F0 cell line. Replacement of tumour cell CM by 10 ng ml-1 basic fibroblast growth factor (bFGF) also results in capillary-like structure formation by BAE cells. An anti-bFGF antibody blocks this effect, showing that bFGF is one of the factors responsible for the angiogenic response induced by B16F10 CM in our assay. Addition of an anti-laminin antibody reduces significantly the formation of capillary-like structures, probably by blocking the attachment of BAE cells to laminin present in Matrigel. The anti-angiogenic compound suramin inhibits in a dose-dependent manner (complete inhibition with 100 microM suramin) the migration and differentiation of BAE cells on type I collagen in response to B16F10 CM. This assay represents a new model system to study tumour-induced angiogenesis in vitro.

Publication types

  • Comparative Study

MeSH terms

  • Adenocarcinoma / pathology
  • Adenocarcinoma / physiopathology*
  • Animals
  • Antibodies / pharmacology
  • Aorta
  • Breast Neoplasms / pathology
  • Breast Neoplasms / physiopathology*
  • Capillaries / cytology*
  • Cattle
  • Cell Differentiation*
  • Cells, Cultured
  • Chemotaxis*
  • Culture Media, Conditioned
  • Endothelium, Vascular / cytology*
  • Fibroblast Growth Factor 2 / pharmacology
  • Humans
  • Laminin / immunology
  • Laminin / pharmacology
  • Melanoma, Experimental / pathology
  • Melanoma, Experimental / physiopathology*
  • Mice
  • Mice, Inbred Strains
  • Neoplasm Invasiveness
  • Neovascularization, Pathologic*
  • Suramin / pharmacology
  • Tumor Cells, Cultured

Substances

  • Antibodies
  • Culture Media, Conditioned
  • Laminin
  • Fibroblast Growth Factor 2
  • Suramin