Selection and characterization of bovine aortic endothelial cells.
This paper reports techniques for isolation, selection and long-term passage of bovine aortic endothelium (BAE). A [3H]thymidine-selection technique was developed to limit overgrowth of cultures by contaminating smooth-muscle cells. The resulting cultures could be passaged for a replicative life span of 35 to 40 doublings and maintained a stable, normal karyotpye throughout this period. Despite the fact that these cultures reached a stable monolayer with density-inhibited growth state, postconfluent cells showed focal areas of a second growth pattern called "sprouting." This was seen only when cultures were maintained at high densities for periods of 1 to 2 weeks. Ultrastructural analysis, as well as immunofluorescence studies with markers for endothelial cells (factor VIII) and smooth-muscle cells (actin), indicates that this phenomenon is not due to overgrowth of a residual population of smooth-muscle cells, but may represent a second growth pattern of the endothelial cells themselves.
D000818:Animals; D001011:Aorta; D002417:Cattle; D002455:Cell Division; D002460:Cell Line; D002469:Cell Separation; D003260:Contact Inhibition; D046508:Culture Techniques; D004727:Endothelium / Q000166:cytology* / Q000648:ultrastructure; D009130:Muscle, Smooth / Q000166:cytology