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J Clin Invest. Nov 1973; 52(11): 2745–2756.
PMCID: PMC302542

Culture of Human Endothelial Cells Derived from Umbilical Veins. IDENTIFICATION BY MORPHOLOGIC AND IMMUNOLOGIC CRITERIA

Abstract

Endothelial cells were isolated from freshly obtained human umbilical cords by collagenase digestion of the interior of the umbilical vein. The cells were grown in tissue culture as a homogeneous population for periods up to 5 mo and some lines were subcultured for 10 serial passages. During the logarithmic phase of cell growth, cell-doubling time was 92 h. Light, phase contrast, and scanning electron microscopy demonstrated that cultured human endothelial cells grew as monolayers of closely opposed, polygonal large cells whereas both cultured human fibroblasts and human smooth muscle cells grew as overlapping layers of parallel arrays of slender, spindle-shaped cells. By transmission electron microscopy, cultured endothelial cells were seen to contain cytoplasmic inclusions (Weibel-Palade bodies) characteristic of in situ endothelial cells. These inclusions were also found in endothelial cells lining umbilical veins but were not seen in smooth muscle cells or fibroblasts in culture or in situ. Cultured endothelial cells contained abundant quantities of smooth muscle actomyosin. Cultured endothelial cells also contained ABH antigens appropriate to the tissue donor's blood type; these antigens were not detectable on cultured smooth muscle cells or fibroblasts. These studies demonstrate that it is possible to culture morphologically and immunologically identifiable human endothelial cells for periods up to 5 mo.

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  • TODD AS. The histological localisation of fibrinolysin activator. J Pathol Bacteriol. 1959 Jul;78:281–283. [PubMed]
  • Spaet TH, Erichson RB. The vascular wall in the pathogenesis of thrombosis. Thromb Diath Haemorrh Suppl. 1966;21:67–86. [PubMed]
  • Baumgartner HR, Tranzer JP, Studer A. An electron microscopic study of platelet thrombus formation in the rabbit with particular regard to 5-hydroxytryptamine release. Thromb Diath Haemorrh. 1967 Dec 31;18(3-4):592–604. [PubMed]
  • Tranzer JP, Baumgartner HR. Filling gaps in the vascular endothelium with blood platelets. Nature. 1967 Dec 16;216(5120):1126–1128. [PubMed]
  • Warren BA, de Bono AH. The ultrastructure of initial stages of platelet aggregation and adhesion to damaged vessel walls in vivo. Br J Exp Pathol. 1970 Aug;51(4):415–422. [PMC free article] [PubMed]
  • Zeldis SM, Nemerson Y, Pitlick FA, Lentz TL. Tissue factor (thromboplastin): localization to plasma membranes by peroxidase-conjugated antibodies. Science. 1972 Feb 18;175(4023):766–768. [PubMed]
  • Stemerman MB, Spaet TH. The subendothelium and thrombogenesis. Bull N Y Acad Med. 1972 Feb;48(2):289–301. [PMC free article] [PubMed]
  • Becker CG, Nachman RL. Contractile proteins of endothelial cells, platelets and smooth muscle. Am J Pathol. 1973 Apr;71(1):1–22. [PMC free article] [PubMed]
  • Gore I, Takada M, Austin J. Ultrastructural basis of experimental thrombocytopenic purpura. Arch Pathol. 1970 Sep;90(3):197–205. [PubMed]
  • Roy AJ, Djerassi I. Effects of platelet transfusions: plug formation and maintenance of vascular integrity. Proc Soc Exp Biol Med. 1972 Jan;139(1):137–142. [PubMed]
  • Strum JM, Junod AF. Radioautographic demonstration of 5-hydroxytryptamine- 3 H uptake by pulmonary endothelial cells. J Cell Biol. 1972 Sep;54(3):456–467. [PMC free article] [PubMed]
  • POMERAT CM, SLICK WC. Isolation and growth of endothelial cells in tissue culture. Nature. 1963 Jun 1;198:859–861. [PubMed]
  • Fryer DG, Birnbaum G, Luttrell CN. Human endothelium in cell culture. J Atheroscler Res. 1966 Mar-Apr;6(2):151–163. [PubMed]
  • Jarmolych J, Daoud AS, Landau J, Fritz KE, McElvene E. Aortic media explants. Cell proliferation and production of mucopolysaccharides, collagen, and elastic tissue. Exp Mol Pathol. 1968 Oct;9(2):171–188. [PubMed]
  • Ross R. The smooth muscle cell. II. Growth of smooth muscle in culture and formation of elastic fibers. J Cell Biol. 1971 Jul;50(1):172–186. [PMC free article] [PubMed]
  • Nachman RL, Marcus AJ, Safier LB. Platelet thrombosthenin: subcellular localization and function. J Clin Invest. 1967 Aug;46(8):1380–1389. [PMC free article] [PubMed]
  • Becker CG, Murphy GE. Demonstration of contractile protein in endothelium and cells of the heart valves, endocardium, intima, arteriosclerotic plaques, and Aschoff bodies of rheumatic heart disease. Am J Pathol. 1969 Apr;55(1):1–37. [PMC free article] [PubMed]
  • Becker CG. Demonstration of actomyosin in mesangial cells of the renal glomerulus. Am J Pathol. 1972 Jan;66(1):97–110. [PMC free article] [PubMed]
  • WEIBEL ER, PALADE GE. NEW CYTOPLASMIC COMPONENTS IN ARTERIAL ENDOTHELIA. J Cell Biol. 1964 Oct;23:101–112. [PMC free article] [PubMed]
  • SZULMAN AE. The histological distribution of blood group substances A and B in man. J Exp Med. 1960 Jun 1;111:785–800. [PMC free article] [PubMed]
  • SZULMAN AE. THE HISTOLOGICAL DISTRIBUTION OF THE BLOOD GROUP SUBSTANCES IN MAN AS DISCLOSED BY IMMUNOFLUORESCENCE. III. THE A, B, AND H ANTIGENS IN EMBRYOS AND FETUSES FROM 18 MM IN LENGTH. J Exp Med. 1964 Apr 1;119:503–516. [PMC free article] [PubMed]
  • Majno G, Shea SM, Leventhal M. Endothelial contraction induced by histamine-type mediators: an electron microscopic study. J Cell Biol. 1969 Sep;42(3):647–672. [PMC free article] [PubMed]
  • Gabbiani G, Hirschel BJ, Ryan GB, Statkov PR, Majno G. Granulation tissue as a contractile organ. A study of structure and function. J Exp Med. 1972 Apr 1;135(4):719–734. [PMC free article] [PubMed]
  • Kemp RB, Jones BM, Gröschel-Stewart U. Aggregative behaviour of embryonic chick cells in the presence of antibodies directed against actomyosins. J Cell Sci. 1971 Jul;9(1):103–122. [PubMed]
  • HOGMAN CF. Blood group antigens A and B determined by means of mixed agglutination on cultured cells of human fetal kidney, liver, spleen, lung, heart and skin. Vox Sang. 1959 Sep;4:319–332. [PubMed]
  • HOLBOROW EJ, BROWN PC, GLYNN LE, HAWES MD, GRESHAM GA, O'BRIEN TF, COOMBS RR. The distribution of the blood group. A antigen in human tissues. Br J Exp Pathol. 1960 Aug;41:430–437. [PMC free article] [PubMed]
  • Davidsohn I, Ni LY. Immunocytology of cancer. Acta Cytol. 1970 May;14(5):276–282. [PubMed]
  • Davidsohn I, Ni LY, Stejskal R. Tissue isoantigens A,B, and H in carcinoma of the stomach. Arch Pathol. 1971 Dec;92(6):456–464. [PubMed]
  • Kovarik S, Davidsohn I, Stejskal R. ABO antigens in cancer. Detection with the mixed cell agglutination reaction. Arch Pathol. 1968 Jul;86(1):12–21. [PubMed]
  • Szulman AE. The A, B and H blood-group antigens in human placenta. N Engl J Med. 1972 May 11;286(19):1028–1031. [PubMed]
  • CHESSIN LN, BRAMSON S, KUHNS WJ, HIRSCHHORN K. STUDIES ON THE A, B, O(H) BLOOD GROUPS ON HUMAN CELLS IN CULTURE. Blood. 1965 Jun;25:944–953. [PubMed]
  • Franks D, Dawson A. Variation in the expression of blood group antigen A in clonal cultures of rabbit cells. Exp Cell Res. 1966 Jun;42(3):543–561. [PubMed]
  • Dawson A, Franks D. Factors affecting the expression of blood group antigen A in cultured cells. Exp Cell Res. 1967 Aug;47(1):377–385. [PubMed]
  • Friedhoff F, Kuhns WJ. Detection and characterization of blood group antigens on untransformed human amnion cells. Transfusion. 1968 Jul-Aug;8(4):244–249. [PubMed]
  • Jaffe EA, Hoyer LW, Nachman RL. Synthesis of antihemophilic factor antigen by cultured human endothelial cells. J Clin Invest. 1973 Nov;52(11):2757–2764. [PMC free article] [PubMed]

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