Modulation of extracellular matrix proteins by endothelial cells undergoing angiogenesis in vitro.

Angiogenesis results in part from the response of endothelial cells to the integrated action of morphogenic factors and extracellular matrix proteins. In this study we identified specific components of the extracellular matrix that were modulated in endothelial cells derived from bovine aorta and rat cerebral microvessels, both of which spontaneously form cords and tubes under standard culture conditions. SPARC (secreted protein, acidic and rich in cysteine) was upregulated 4.2-fold in aortic and 10-fold in microvascular cultures that had organized into cords and/or tubes. This Ca(2+)-binding glycoprotein was synthesized primarily by endothelial cells in the process of cord formation. Transcription of type I collagen was initiated in aortic endothelial cells undergoing angiogenesis in vitro and showed a 12-fold increase in similar cultures of microvascular cells. Type VIII collagen protein was upregulated to a lesser degree (4.3-fold in aortic and 1.8-fold in microvascular cells). Dense cytoplasmic staining for these two collagen types was seen in cells directly participating in the organization of cords. In contrast, the disparate levels of fibronectin observed in both types of endothelium indicated an indirect or secondary role for this glycoprotein in cord/tube formation in vitro. These results identify SPARC, type I collagen, and type VIII collagen as extracellular matrix components that are actively synthesized by endothelial cells undergoing angiogenesis in vitro. Moreover, expression of these proteins during the formation of tubes and cords appears to follow a biosynthetic program that is common to endothelial cells from both the macrovasculature and microvasculature.