Microvascular Endothelial Cells and Their Relationship to Angiogenesis
Angeles, Susan M.
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Angiogenesis is the formation of new blood vessels, a process in which endothelial cells play a major role. In this project, it was determined that endothelial cells, along with a mixed population of cells, could be harvested from angiogenesis chambers (developed by Dvorak et aI., 1987) implanted in Sprague Dawley rats. Endothelial cells were evidenced by their uptake of DiI-Ac-LDL, an endothelial cell marker. It was found that endothelial cells could be obtained when the extracted fibrin gels were left whole, resting on a gelatin substrate (to which the endothelial cells could better attach themselves) in Endothelial Basal Medium supplemented with 10% fetal bovine serum, antibiotics gentamicin/amphotericin B, and 10 ng/ml basic fibroblast growth factor (bFGF). However, sustained endothelial cell growth was poor. An isolation procedure for endothelial cells was performed, but did not yield endothelial cells, demonstrated by the cells' lack of DiI-Ac-LDL uptake. In addition, the anti-proliferative effects of suramin, an antitrypanosomal drug (Stein et ai., 1989), and/or steroids (angiostatic U-24067 and non-angiostatic testosterone) were examined using two different endothelial cell lines, bovine adrenal capillary (BACE) and transformed mouse cerebral microvessel (TEn) endothelial cells. Greatest endothelial cell inhibition was observed with suramin alone and steroid alone (U-24067 or testosterone) at their highest respective doses tested. Suramin and angiostatic steroid U-24067 did not yield the expected synergistic inhibition of BACE and TEn cell growth as was shown in the chorioallantoic membrane (CAM) assay on tumor growth (Wilks et al., 1990). A synergistic inhibitory effect, however, was observed with suramin and testosterone in this preliminary experiment. From the dose response experiments, it was concluded that the anti-proliferative effects of suramin and steroid were non-specific for angiostatic steroids since no synergistic inhibitory effect was observed with suramin and U -24067 and since greatest cell inhibition was observed with suramin alone and steroid alone regardless of whether the steroid was angiostatic or not.