Reorganization of endothelial cord-like structures on basement membrane complex (Matrigel): involvement of transforming growth factor beta 1.
The formation of capillary-like network structures by cultured vascular endothelial cells on reconstituted basement membrane matrix, Matrigel, models endothelial cell differentiation, the final step of angiogenesis (Kubota et al., 1988; Grant et al., 1989). When endothelial cells derived from bovine aorta and brain capillaries were plated on Matrigel, DNA synthesis was suppressed and a network of capillary-like structures rapidly formed in 8-12 h. With time, the network broke down, resulting in dense cellular cords radiating from multiple cellular clusters in 16-24 h. Finally, multicellular aggregates of cells were formed as the network underwent further retraction. Network regression was prevented when either dithiothreitol (DTT) or anti-TGF-beta 1 antibodies were added during the assay. The addition of exogenous TGF-beta 1 promoted the regression of endothelial cells into the clusters. This response to TGF-beta 1 was blocked by potent serine threonine protein kinase inhibitors, H-7 and HA100. TGF-beta 1 was released from polymerized Matrigel by incubation with Dulbecco's modified eagle's medium (DMEM) in the absence of cells. The Matrigel-conditioned DMEM inhibited endothelial DNA synthesis even in the presence of anti-TGF-beta 1 antibodies. These results suggest that TGF-beta 1 and possibly other soluble factors from Matrigel may be important for differentiation and remodeling of endothelial cells in a capillary network with possible implications for wound healing and development.
Laboratory of Cardiovascular Science, National Institute on Aging, Baltimore, Maryland 21224.
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Kuzuya
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JL
Kinsella
D000818:Animals; D002196:Capillaries / Q000033:anatomy & histology; D002417:Cattle; D002478:Cells, Cultured; D003094:Collagen / Q000494:pharmacology*; D004247:DNA / Q000037:antagonists & inhibitors / Q000096:biosynthesis; D004338:Drug Combinations; D004730:Endothelium, Vascular / Q000166:cytology / Q000187:drug effects* / Q000502:physiology*; D007797:Laminin / Q000494:pharmacology*; D047428:Protein Kinase Inhibitors; D011509:Proteoglycans / Q000494:pharmacology*; D016212:Transforming Growth Factor beta / Q000502:physiology*
D004338:Drug Combinations; D007797:Laminin; D047428:Protein Kinase Inhibitors; D011509:Proteoglycans; D016212:Transforming Growth Factor beta; C059485:matrigel; D003094:Collagen; D004247:DNA