Endothelial cell differentiation into capillary-like structures in response to tumour cell conditioned medium: a modified chemotaxis chamber assay.
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.
Garrido
T
T
Pharmacia Antibióticos Farma SA, Research Department, Madrid, Spain.
Riese
H H
HH
Aracil
M
M
Pérez-Aranda
A
A
D000230:Adenocarcinoma / Q000473:pathology / Q000503:physiopathology*; D000818:Animals; D000906:Antibodies / Q000494:pharmacology; D001011:Aorta; D001943:Breast Neoplasms / Q000473:pathology / Q000503:physiopathology*; D002196:Capillaries / Q000166:cytology*; D002417:Cattle; D002454:Cell Differentiation; D002478:Cells, Cultured; D002633:Chemotaxis; D017077:Culture Media, Conditioned; D004730:Endothelium, Vascular / Q000166:cytology*; D016222:Fibroblast Growth Factor 2 / Q000494:pharmacology; D006801:Humans; D007797:Laminin / Q000276:immunology / Q000494:pharmacology; D008546:Melanoma, Experimental / Q000473:pathology / Q000503:physiopathology*; D051379:Mice; D008815:Mice, Inbred Strains; D009361:Neoplasm Invasiveness; D009389:Neovascularization, Pathologic; D013498:Suramin / Q000494:pharmacology; D014407:Tumor Cells, Cultured
D000906:Antibodies; D017077:Culture Media, Conditioned; D007797:Laminin; D016222:Fibroblast Growth Factor 2; D013498:Suramin