HUMAN ENDOTHELIAL-CELL MORPHOLOGY AND AUTACOID EXPRESSION

Human umbilical vein endothelial (HUVE) cells plated on plastic or gel atin-coated dishes grow as a ''cobblestone'' monolayer. By contrast, e ndothelial cells cultured on a complex matrix (e.g., Matrigel) form th ree-dimensional, capillary-like structures. In the current study, we v erified the capillary phenotype of the latter structures and asked whe ther the morphological changes induced by extracellular matrix also af fect human endothelial gene expression and function in vitro. Concentr ations of cellular fibronectin, prostacyclin, and endothelin-1 were me asured in the conditioned media by enzyme-linked immunosorbent and rad ioimmunoassays. Steady-state concentrations of HUVE mRNA were estimate d by reverse transcription-polymerase chain reaction and quantified by Northern analyses to assess fibronectin and endothelin-1 gene express ion. We found that the subjacent extracellular matrix affects the morp hology, proliferation, and differentiation of HUVE cells in vitro. Cel ls cultured on gelatin were more mitotically active, expressed signifi cantly less cellular fibronectin, made similar amounts of prostacyclin , and secreted significantly more endothelin-1 compared with the same cells grown on a Matrigel substrate.