Macro- and microvascular endothelial cells in vitro: maintenance of biochemical heterogeneity despite loss of ultrastructural characteristics.
Microvascular endothelial cells from bovine adrenal medulla and brain and macrovessel endothelial cells from bovine aorta were isolated and cultured under similar conditions in order to determine morphologic and biochemical heterogeneity in vitro. All three cell types exhibited nearly identical ultrastructural morphology and two-dimensional gel protein patterns of 35S-methionine-labeled whole cells. Two-dimensional gel analysis of 35S-methionine-labeled plasma membrane proteins however, revealed two-dimensional gel protein patterns unique to the tissue type from which the endothelial cells were isolated. This suggests that the functional significance of these specific endothelial cell types is manifested primarily in surface-associated proteins and that many of the differences are sustained in culture. To determine the potential of aorta, brain, and adrenal medulla endothelial cell (EC) cultures to respond to developmentally significant signals, morphology, growth pattern, and cell surface proteins were monitored in the presence and absence of growth factors. A 17 to 26% increase in cell density as well as an increase in the number of elongated and overlapping cells resulted when all three EC types were exposed to a mitogenic medium. Additionally, expression of specific glycoprotein profiles, as determined by Concanavalin A Western blotting of two-dimensional gels, was dependent on the presence or absence of growth factors in the medium. The ability to induce this morphologic and biochemical variation in the three endothelial cell types was maintained into later passage. Taken together, these data imply that endothelial cells isolated from different tissues exhibit and maintain biochemical heterogeneity and do not completely dedifferentiate into a common endothelial cell type in culture. Furthermore, expression of specific subsets of cell surface proteins is dependent on environmental conditions, and in some cases is both cell-type and media-type dependent. Thus, even though endothelial cells are considered terminally differentiated cells, there exists additional or "latent" heterogeneity in the ability of these different cells to respond to "developmental signals" (i.e. mitogenic medium) in vitro.
Stolz
D B
DB
Program in Molecular and Cellular Biology, University of Massachusetts, Amherst 01003.
Jacobson
B S
BS
D000313:Adrenal Medulla / Q000098:blood supply / Q000166:cytology; D000818:Animals; D001011:Aorta / Q000166:cytology; D015153:Blotting, Western; D001921:Brain / Q000098:blood supply / Q000166:cytology; D002417:Cattle; D002462:Cell Membrane / Q000737:chemistry / Q000648:ultrastructure; D002469:Cell Separation; D002478:Cells, Cultured; D015180:Electrophoresis, Gel, Two-Dimensional; D004730:Endothelium, Vascular / Q000737:chemistry* / Q000648:ultrastructure; D006023:Glycoproteins / Q000032:analysis / Q000378:metabolism; D008565:Membrane Proteins / Q000032:analysis / Q000378:metabolism; D008715:Methionine / Q000378:metabolism; D008833:Microcirculation; D008854:Microscopy, Electron; D013462:Sulfur Radioisotopes
D006023:Glycoproteins; D008565:Membrane Proteins; D013462:Sulfur Radioisotopes; D008715:Methionine