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 karyotype 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.
Key wordsthymidine selection endothelium contact inhibition replicative life span
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- 1.Gimbrone, M. A. 1976. Culture of vascular endothelium. In: Theodore H. Spaet (Ed.),Progress in Hemostasis and Thrombosis. Vol. 3. Grune and Stratton. New York, pp. 1–28.Google Scholar
- 6.Majno, G. 1965. Ultrastructure of the vascular membrane. In:Handbook of Physiology. Vol. III. American Physiological Society. Washington, pp. 2293–2397.Google Scholar
- 12.Harnden, D. G. 1974. Skin culture and solid tumor technique In: Jorge, J. Yunis (Ed.),Human Chromosome Methodology. Academic Press, New York, pp. 167–184.Google Scholar
- 18.Gospodarowicz, D., and A. L. Mecher. Control of cellular proliferation by the fibroblast and epiderma growth factors: Nat. Cancer Inst. Monogr. 48, in press.Google Scholar
- 20.Tilney, L. G. 1975. The role of actin in non-muscle cell motility. In: S. Inone, and R. E. Stephens (Eds.),Molecules and Cell Movement. Raven, New York, pp. 339–388.Google Scholar
- 23.Rutherford, R. B., and E. P. Benditt. 1976. Clustering of replicating cells in the aortic endothelium. Proc. Natl. Acad. Sci. U.S.A. 73: 652–653.Google Scholar