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Adhesion Molecules on Isolated and Cultured Microvascular Endothelial Cells Demonstrated by Immunofluorescence and Immune Electron Microscopy

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Part of the book series: Methods in Molecular Biology™ ((MIMB,volume 75))

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Abstract

In vitro systems of cultured endothelial cells (EC) represent appropriate tools for simulating interactions of cells involved in leucocyte homing or in tumor invasion. Such processes require the adhesion of cooperating cells via receptors classified as adhesion molecules (AM). EC equipped with AM are able to interact with other cells that bear counter-receptors (cell-cell communications) or with molecules of the extracellular matrix (ECM; cell-matrix communications). AM are routinely demonstrated in cell cultures or tissue sections by immunofluorescence or immunohistochemistry at the light- or electron-microscopical level.

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References

  1. Spicer, S. S. and Schulte, B. A. (1992) Diversity of cell glycoconjugates shown histochemically: a perspective. J. Histochem. Cytochem. 40, 1–38.

    CAS  PubMed  Google Scholar 

  2. Lee, Y. C. and Lee, R. T. (1995) Carbohydrate-protein interactions: basis of glycobiology. Act. Chem. Res. 28, 321–327.

    CAS  Google Scholar 

  3. Wu, J. T. (1993) Advanced glycosylation end products: a new disease marker for diabetes and aging. J. Clin. Lab. Anal. 7, 252–255.

    CAS  PubMed  Google Scholar 

  4. Jaffe, E. A. (1987) Cell biology of endothelial cells. Hum. Pathol. 18, 234–239.

    CAS  PubMed  Google Scholar 

  5. Augustin, H. G., Kozian, D. H., and Johnson, R. C. (1994) Differentiation of endothelial cells: analysis of the constitutive and activated endothelial cell phenotypes. BioEssays 16, 901–906.

    CAS  PubMed  Google Scholar 

  6. Hormia, M. and Virtanen, I. (1986) Endothelium—an organized monolayer of highly specialized cells. Med. Biol. 64, 247–266.

    CAS  PubMed  Google Scholar 

  7. Risau, W. (1995) Differentiation of endothelium. FASEB J. 9, 926–933.

    CAS  PubMed  Google Scholar 

  8. Duijvestijn, A. M., Rep, M., Hendriks, H. R., and Kraal, G. (1990) Functional capacities of high endothelial venules appear not to be controlled by recirculating lymphocytes. Immunobiology 180, 295–307.

    CAS  PubMed  Google Scholar 

  9. Kraal, G., Duijvestijn, A. M., and Hendriks, H. H. (1987) The endothelium of the high endothelial venule: a specialized endothelium with unique properties. Exp. Cell Biol. 55, 1–10.

    CAS  PubMed  Google Scholar 

  10. Jalkanen, S., Steere, A. C., Fox, R. I., and Butcher, E. C. (1986) A distinct endothelial cell recognition system that controls lymphocyte traffic into inflamed synovium. Science 233, 556–559.

    CAS  PubMed  Google Scholar 

  11. Freemont, A. J. (1988) Functional and biosynthetic changes in endothelial cells of vessels in chronically inflamed tissues: evidence for endothelial control of lymphocyte entry into diseased tissues. J. Pathol. 155, 225–230.

    CAS  PubMed  Google Scholar 

  12. Ziff, M. (1989) Role of endothelium in chronic inflammation. Springer Semin. Immunopathol. 11, 199–214.

    CAS  PubMed  Google Scholar 

  13. Springer, T. A. (1990) Adhesion receptors of the immune system. Nature 346, 425–434.

    CAS  PubMed  Google Scholar 

  14. Boehncke, W.-H., Kellner, I., Konter, U., and Sterry, W. (1992) Differential expression of adhesion molecules on infiltrating cells in inflammatory dermatoses. J. Am. Acad. Dermatol. 26, 907–913.

    CAS  PubMed  Google Scholar 

  15. Smith, C. W. (1993) Endothelial adhesion molecules and their role in inflammation. Can. J. Physiol. Pharmacol. 71, 76–87.

    CAS  PubMed  Google Scholar 

  16. Williams, T. J. and Hellewell, P. G. (1992) Endothelial cell biology. Adhesion molecules involved in the microvascular inflammatory response. Am. Rev. Respir. Dis. 146, S45–S50.

    CAS  PubMed  Google Scholar 

  17. Jutila, M. A. (1992) Leukocyte traffic to sites of inflammation. APMIS 100, 191–201.

    CAS  PubMed  Google Scholar 

  18. Vachula M. and Van Epps, D. E. (1992) In vitro models of lymphocyte transendothelial migration. Invasion Metastasis 12, 66–81.

    CAS  PubMed  Google Scholar 

  19. Hamann, A. (1992) Mechanisms of lymphocyte traffic and cell targeting. Int. J. Cancer 7(Suppl.), 19–23.

    CAS  Google Scholar 

  20. Swerlick, R. A. and Lawley, T. J. (1993) Role of microvascular endothelial cells in inflammation. J. Invest. Dermatol. 100, 111S–115S.

    CAS  PubMed  Google Scholar 

  21. Cronstein, B. N. and Weissmann, G. (1993) The adhesion molecules of inflammation. Arthritis Rheum. 36, 147–157.

    CAS  PubMed  Google Scholar 

  22. Stad, R. K. and Buurman, W. A. (1994) Current views on structure and function of endothelial adhesion molecules. Cell Adh. Commun. 2, 261–268.

    CAS  Google Scholar 

  23. Dejana, E., Breviario, F., and Caveda, L. (1994) Leucocyte-endothelial cell adhesive receptors. Clin. Exp. Rheumatol. 12(Supp1. 10), S25–S28.

    PubMed  Google Scholar 

  24. Adams, D. H. and Shaw, S. (1994) Leucocyte-endothelial interactions and regulation of leucocyte migration. Lancet 343, 831–836.

    CAS  PubMed  Google Scholar 

  25. Bischoff, J. (1995) Approaches to studying cell adhesion molecules in angiogenesis. Trends Cell Biol. 5, 60–65.

    Google Scholar 

  26. Cartwright, J. E., Whitley, G. St. J., and Johnstone, A. P. (1995) The expression and release of adhesion molecules by human endothelial cell lines and their consequent binding of lymphocytes. Exp. Cell Res. 217, 329–335.

    CAS  PubMed  Google Scholar 

  27. Gimbrone, M. A., Jr., Cotran, R. S., and Folkman, J. (1974) Human vascular endothelial cells in culture. J. Cell Biol. 60, 673–484.

    CAS  PubMed  Google Scholar 

  28. Moyer, C. F., Dennis, P. A., Majno, G., and Joris, I. (1988) Venular endothelium in vitro: isolation and characterization. In Vitro Cell. Dev. Biol. 24, 359–368.

    CAS  PubMed  Google Scholar 

  29. Jaffe, E. A., Nachman, R. L., Becker, C. G., and Minick, C. R. (1973) Culture of human endothelial cells derived from umbilical veins. Identification by morphologic and immunologic criteria. J. Clin. Invest. 52, 2745–2756.

    CAS  PubMed Central  PubMed  Google Scholar 

  30. Weinstein, R. and Wenc, K. (1986) Growth factor responses of human arterial endothelial cells in vitro. In Vitro Cell. Dev. Biol. 22, 549–556.

    CAS  PubMed  Google Scholar 

  31. Introna, M., Colotta, F., Sozzani, S., Dejana, E., and Mantovani, A. (1994) Pro-and anti-inflammatory cytokines interactions with vascular endothelium. Clin. Exp. Rheumatol. 12(Supp1. 10), S19–S23.

    PubMed  Google Scholar 

  32. Schiffrin, E. L. (1994) The endothelium and control of blood vessel function in health and disease. Clin. Invest. Med. 17, 602–620.

    CAS  PubMed  Google Scholar 

  33. Roberts, W. G. and Palade, G. E. (1995) Increased microvascular permeability and endothelial fenestration induced by vascular endothelial growth factor. J. Cell Sci. 108, 2369–2379.

    CAS  PubMed  Google Scholar 

  34. McMillen, M. A. and Sumpio, B. E. (1995) Endothelins: polyfunctional cytokines. J. Am. Coll. Surg. 180, 621–637.

    CAS  PubMed  Google Scholar 

  35. Yao, J., Bone, R. C., and Sawnhey, R. S. (1995) Differential effects of tumor necrosis factor-alpha on the expression of fibronectin and collagen genes in cultured bovine endothelial cells. Cell. Mol. Biol. Res. 41, 17–28.

    CAS  PubMed  Google Scholar 

  36. Chuluyan, H. E., Schall, T. J., Yoshimura, T., and Issekutz, A. C. (1995) IL-1 activation of endothelium supports VLA-4 (CD49d/CD29)-mediated monocyte transendothelial migration to C5a, MIP-1α, RANTES, and PAF but inhibits migration to MCP-1: a regulatory role for endothelium-derived MCP-1. J. Leukocyte Biol. 58, 71–79.

    CAS  PubMed  Google Scholar 

  37. Ristimaki, A. and Viinikka, L. (1992) Modulation of prostacyclin production by cytokines in vascular endothelial cells. Prostagland Leukotrienes Essential Fatty Acids 47, 93–99.

    CAS  Google Scholar 

  38. Kramer, R. H., Bensch, K. G., Davison, P. M., and Karasek, M. A. (1984) Basal lamina formation by cultured microvascular endothelial cells. J. Cell Biol. 99, 692–698.

    CAS  PubMed  Google Scholar 

  39. Bowersox, J. C. and Sorgente, N. (1987) Differential effects of soluble and immobilized fibronectins on aortic endothelial cell proliferation and attachment. In Vitro Cell. Dev. Biol. 23, 759–764.

    CAS  PubMed  Google Scholar 

  40. Nicosia, R. F. and Madri, J. A. (1987) The microvascular extracellular matrix. Am. J. Pathol. 128, 78–90.

    CAS  PubMed  Google Scholar 

  41. Lou, D.-A. and Hu, F. (1987) Co-distribution of von Willebrand factor and fibronectin in cultured rhesus endothelial cells. Histochem. J. 19, 431–438.

    CAS  PubMed  Google Scholar 

  42. Kawasaki, S., Mori, M., and Awai, M. (1989) Capillary growth of rat aortic segments cultured in collagen gel without serum. Acta Pathol. Jpn. 39, 712–718.

    CAS  PubMed  Google Scholar 

  43. Tijburg, P. N. M., Ryan, J., Stem, D. M., Wollitzky, B., Rimon, S., Rimon, A., Handley, D., Nawroth, P., Sixma, J. J., and de Groot, P. G. (1991) Activation of the coagulation mechanism on tumor necrosis factor-stimulated cultured endothelial cells and their extracellular matrix. J. Biol. Chem. 266, 12,067–12,074.

    CAS  PubMed  Google Scholar 

  44. Fournier, N. and Doillon, C. J. (1992) In vitro angiogenesis in fibrin matrices containing fibronectin or hyaluronic acid. Cell Biol. Int. Rep. 16, 1251–1263.

    CAS  PubMed  Google Scholar 

  45. Berge, V., Johnson, E., Høgåsen, K., and Hetland, G. (1992) Human umbilical vein endothelial cells synthesize S-protein (vitronectin) in vitro. Scand. J. Immunol. 36, 119–123.

    CAS  PubMed  Google Scholar 

  46. Fenyves, A. M., Behrens, J., and Spanel-Borowski, K. (1993) Cultured microvascular endothelial cells (MVEC) differ in cytoskeleton, expression of cadherins and fibronectin matrix. A study under the influence of interferon-γ. J. Cell Sci. 106, 879–890.

    CAS  PubMed  Google Scholar 

  47. Schnaper, H. W., Kleinman, H. K., and Grant, D. S. (1993) Role of laminin in endothelial cell recognition and differentiation. Kidney Int. 43, 20–25.

    CAS  PubMed  Google Scholar 

  48. Inagaki, M., Baxter, B. T., Cisler, J., Davis, V., Prorok, G. D., and Langnas, A. N. (1993) Synthesis of interstitial collagen by hepatic sinusoidal endothelial cells, Cells of the Hepatic Sinusoid, vol 4. (Knook, D. L., and Wisse, E., eds.), Kupffer Cell Foundation, Leiden, The Netherlands, pp. 268–270.

    Google Scholar 

  49. Ziats, N. P. and Anderson, J. M. (1993) Human vascular endothelial cell attachment and growth inhibition by type V collagen. J. Vasc. Surg. 17, 710–718.

    CAS  PubMed  Google Scholar 

  50. Nicosia, R. F. and Tuszynski, G. P. (1994) Matrix-bound thrombospondin promotes angiogenesis in vitro. J. Cell Biol. 124, 183–193.

    CAS  PubMed  Google Scholar 

  51. Fournier, N. and Doillon, C. J. (1994) In vitro effects of extracellular matrix and growth factors on endothelial cell migration and vessel formation. Cells Materials 4, 399–408.

    CAS  Google Scholar 

  52. Sage, E. H. and Vernon, R. B. (1994) Regulation of angiogenesis by extracellular matrix: the growth and the glue. J. Hypert. 12(Suppl. 10), S145–S152.

    CAS  Google Scholar 

  53. Wu, C., Chung, A. E., and McDonald, J. A. (1995) A novel role for α3β1 integrins in extracellular matrix assembly. J. Cell Sci. 108, 2511–2523.

    CAS  PubMed  Google Scholar 

  54. Qi, J. and Kreutzer, D. L. (1995) Fibrin activation of vascular endothelial cells. Induction of IL-8 expression. J. Immunol. 155, 867–876.

    CAS  PubMed  Google Scholar 

  55. Passaniti, A., Taylor, R. M., Pili, R., Guo, Y., Long, P. V., Haney, J. A., Pauly, R. R., Grant, D. S., and Martin, G. R. (1992) Methods in laboratory investigation. A sample, quantitative method for assessing angiogenesis and antiangiogenic agents using reconstituted basement membrane, heparin, and fibroblast growth factor. Lab. Invest. 67, 519–528.

    CAS  PubMed  Google Scholar 

  56. Vinters, H. V., Reave, S., Costello, P., Girvin, J. P., and Moore, S. A. (1987) Isolation and culture of cells derived from human cerebral microvessels. Cell Tissue Res. 249, 657–667.

    CAS  PubMed  Google Scholar 

  57. Coutinho, G. C., Durieu-Trautmann, O., Strosberg, A. D., and Couraud, P. O. (1991) Catecholamines stimulate the IFN-γ-induced class II MHC expression on bovine brain capillary endothelial cells. J. Immunol. 147, 2525–2529.

    CAS  PubMed  Google Scholar 

  58. Dorovini-Zis, K., Bowman, P. D., and Prameya, R. (1992) Adhesion and migration of human polymorphonuclear leukocytes across cultured bovine brain microvessel endothelial cells. J. Neuropathol. Exp. Neurol. 51, 194–205.

    CAS  PubMed  Google Scholar 

  59. Vorbrodt, A. W., and Trowbridge, R. S. (1991) Ultracytochemical characteristics of cultured sheep brain microvascular endothelial cells. J. Histochem. Cytochem. 39, 1555–1563.

    CAS  PubMed  Google Scholar 

  60. Dropulic, B. and Masters, C. L. (1987) Culture of mouse brain capillary endothelial cell lines that express factor VIII, γ-glutamyl transpeptidase, and form juctional complexes in vitro. In Vitro Cell. Dev. Biol. 23, 775–781.

    CAS  PubMed  Google Scholar 

  61. Canfield, A. E. and Schor, A. M. (1994) Heterogeneity in collagen biosynthesis by sprouting retinal endothelial cells. J. Cell. Physiol. 159, 19–28.

    CAS  PubMed  Google Scholar 

  62. Rymaszewski, Z., Szymanski, P. T., Abplanalp, W. A., Myatt, L., Di Salvo, J., and Cohen, R. M. (1992) Human retinal vascular cells differ from umblilical cells in synthetic functions and their response to glucose. PSEBM 199, 183–191.

    CAS  Google Scholar 

  63. Su, T. and Gillies, M. C. (1992) A simple method for the in vitro culture of human retinal capillary endothelial cells. Invest. Ophthalmol. Vis. Sci. 33, 2809–2813.

    CAS  PubMed  Google Scholar 

  64. Hanneken, A., Lutty, G. A., McLeod, D. S., Robey, F., Harvey, A. K., and Hjelmeland, L. M. (1989) Localization of basic fibroblast growth factor to the developing capillaries of the bovine retina. J. Cell. Physiol. 138, 115–120.

    CAS  PubMed  Google Scholar 

  65. Kraling, B. M., Jimenez, S. A., Sorger, T., and Maul, G. G. (1994) Isolation and characterization of microvascular endothelial cells from the adult human dermis and from skin biopsies of patients with systemic sclerosis. Lab. Invest. 71, 745–754.

    CAS  PubMed  Google Scholar 

  66. Hewett, P. W. and Murray, J. C. (1993) Human lung microvessel endothelial cells isolation, culture, and characterization. Microvasc. Res. 46, 89–102.

    CAS  PubMed  Google Scholar 

  67. Magee, J. C., Stone, A. E., Oldham, K. T., and Guice, K. S. (1994) Isolation, culture, and characterization of rat lung microvascular endothelial cells. Am. J. Physiol. 267 (Lung Cell Mol. Physiol 11), L433–L441.

    CAS  PubMed  Google Scholar 

  68. Gumkowski, F., Kaminska, G., Kaminski, M., Morrissey, L. W., and Auerbach, R. (1987) Heterogeneity of mouse vascular endothelium. Blood Vessels 24, 11–23.

    CAS  PubMed  Google Scholar 

  69. Haraldsen, G., Rugtveit, J., Kvale, D., Scholz, T., Muller, W. A., Hovig, T., and Brandtzaeg, P. (1995) Isolation and longterm culture of human intestinal microvascular endothelial cells. Gut 37, 225–234.

    CAS  PubMed  Google Scholar 

  70. Simionescu, M., and Simionescu, N. (1978) Isolation and characterization of endothelial cells from the heart microvasculature. Microvasc. Res. 16, 426–452.

    CAS  PubMed  Google Scholar 

  71. Grafe, M., Auch-Schwelk, W., Graf, K., Terbeek, D., Hertel, H., Unkelbach, M., Hildebrandt, A., and Fleck, E. (1994) Isolation and characterization of macrovascular and microvascular endothelial cells from human hearts. Am. J. Physiol. 267 (Heart Circ. Physiol. 36), H2138–H2148.

    CAS  PubMed  Google Scholar 

  72. Nishida, M., Carley, W. W., Gerritsen, M. E., Ellingsen, O., Kelly, R. A., and Smith, T. W. (1993) Isolation and characterization of human and rat cardiac microvascular endothelial cells. Am. J. Physiol. 264 (Heart Circ. Physiol. 33), H639–H652.

    CAS  PubMed  Google Scholar 

  73. Gomez, D. E., Hartzler, J. L., Corbitt, R. H., Nason, A. M., and Thorgeirsson, U. P. (1993) Immunomagnetic separation as a final purification step of liver endothelial cells. In Vitro Cell. Dev. Biol. 29A, 451–455.

    CAS  Google Scholar 

  74. Spanel-Borowski, K. (1991) Diversity of ultrastructure in different phenotypes of cultured microvessel endothelial cells isolated from bovine corpus luteum. Cell Tissue Res. 266, 37–49.

    CAS  PubMed  Google Scholar 

  75. Spanel-Borowski, K. and van der Bosch, J. (1990) Different phenotypes of cultured microvessel endothelial cells obtained from bovine corpus luteum. Cell Tissue Res. 261, 35–47.

    CAS  PubMed  Google Scholar 

  76. Miller, R. R. and Rydell, P. A. (1993) Primary culture of microvascular endothelial cells from canine meniscus. J. Orthop. Res. 11, 907–911.

    CAS  PubMed  Google Scholar 

  77. Abbot, S. E., Kaul, A., Stevens, C. R., and Blake, D. R. (1992) Isolation and culture of synovial microvascular endothelial cells. Arthritis Rheum. 35, 401–406.

    CAS  PubMed  Google Scholar 

  78. Scott, P. A. E. and Bucknell, R. (1993) The isolation and culture of microvascular endothelium. J. Cell. Sci. 105, 269–273.

    PubMed  Google Scholar 

  79. Karasek, M. A. (1989) Microvascular endothelial cell culture. J. Invest. Dermatol. 93, 33S–38S.

    CAS  PubMed  Google Scholar 

  80. Hewett, P. W. and Murray, J. C. (1993) Immunomagnetic purification of human microvessel endothelial cells using Dynabeads coated with monoclonal antibodies to PECAM-1. Eur. J. Cell. Biol. 62, 451–454.

    CAS  PubMed  Google Scholar 

  81. Jackson, C. J., Garbett, P. K., Nissen, B., and Schrieber, L. (1990) Binding of human endothelium to Ulex europaeus I-coated Dynabeads: application to the isolation of microvascular endothelium. J. Cell Sci. 96, 257–262.

    PubMed  Google Scholar 

  82. Hynes, R. O. and Lander, A. D. (1992) Contact and adhesive specificities in the associations, migrations, and targeting of cells and axons. Cell 68, 303–322.

    CAS  PubMed  Google Scholar 

  83. Ruoslahti, E., Noble, N. A., Kagami, S., and Border, W. A. (1994) Integrins. Kidney Int. 45(Suppl. 44), S-17–S-22.

    Google Scholar 

  84. Sonnenberg, A. (1992) Laminin receptors in the integrin family. Pathol. Biol. 40, 773–778.

    CAS  PubMed  Google Scholar 

  85. Fan, S.-T. and Edgington, T. S. (1993) Integrin regulation of leukocyte inflammatory functions. J. Immunol. 150, 2972–2980.

    CAS  PubMed  Google Scholar 

  86. Hogg, N. and Berlin, C. (1995) Structure and function of adhesion receptors in leukocyte trafficking. Immunol. Today 16, 327–330.

    CAS  PubMed  Google Scholar 

  87. Cepek, K. L., Shaw, S. K., Parker, C. M., Russell, G. J., Morrow, J. S., Rimm, D. L., and Brenner, M. B. (1994) Adhesion between epithelial cells and T lymphocytes mediated by E-cadherin and the αEβ7 integrin. Nature 372, 190–193.

    CAS  PubMed  Google Scholar 

  88. Lasky, L. A. (1992) Selectins: interpreters of cell-specific carbohydrate information during inflammation. Science 258, 964–969.

    CAS  PubMed  Google Scholar 

  89. Cummings, R. D. and Smith, D. F. (1992) The selectin family of carbohydrate-binding proteins: structure and importance of carbohydrate ligands for cell adhesion. BoEssays 14, 849–856.

    CAS  Google Scholar 

  90. Bevilacqua, M. P., and Nelson, R. M. (1993) Selectins. J. Clin. Invest. 91, 379–387.

    CAS  PubMed Central  PubMed  Google Scholar 

  91. Varki, A. (1994) Selectin ligands. Proc. Natl. Acad. Sci. USA 91, 7390–7397.

    CAS  PubMed  Google Scholar 

  92. Tedder, T. F., Steeber, D. A., Chen, A., and Engel, P. (1995) The selectins vascular adhesion molecules. FASEB J. 9, 866–873.

    CAS  PubMed  Google Scholar 

  93. Briskin, M. J., McEvoy, L. M., and Butcher, E. C. (1993) MAdCAM-1 has homology to immunoglobulin and mucin-like adhesion receptors and to IgA1. Nature 363, 461–464.

    CAS  PubMed  Google Scholar 

  94. Berg, E. L., McEvoy, L. M., Berlin, C., Bargatze, R. F., and Butcher, E. C. (1993) L-selectin-mediated lymphocyte rolling on MAdCAM-1. Nature 366, 695–698.

    CAS  PubMed  Google Scholar 

  95. Sako, D., Chang, X.-J., Barone, K. M., Vachino, G., White, H. M., Shaw, G., Veldman, G. M., Bean, K. M., Ahern, T. J., Furie, B., Cumming, C. A., and Larsen, G. R. (1993) Expression cloning of a functional glycoprotein ligand for P-selectin. Cell 75, 1179–1186.

    CAS  PubMed  Google Scholar 

  96. Lasky, L. A., Singer, M. S., Dowbenko, D., Imai, Y., Henzel, W. J., Grimley, C., Fennie, C., Gillett, N., Watson, S. R., and Rosen, S. D. (1992) An endothelial ligand for L-selectin is a novel mucin-like molecule. Cell 69, 927–938.

    CAS  PubMed  Google Scholar 

  97. Takeichi, M. (1991) Cadherin cell adhesion receptors as a morphogenetic regulator. Science 251, 1451–1455.

    CAS  PubMed  Google Scholar 

  98. Salomon, D., Ayalon, O., Patel-King, R., Hynes, R. O., and Geiger, B. (1992) Extrajunctional distribution of N-cadherin in cultured human endothelial cells. J. Cell Sci. 102, 7–17.

    CAS  PubMed  Google Scholar 

  99. Salmi, M. and Jalkanen, S. (1992) A 90-kilodalton endothelial cell molecule mediating lymphocyte binding in human. Science 257, 1407–1409.

    CAS  PubMed  Google Scholar 

  100. Airas, L., Salmi, M., and Jalkanen, S. (1993) Lymphocyte-vascular adhesion protein-2 is a novel 70-kilodalton molecule in volved in lymphocyte adhesion to vascular endothelium. J. Immunol. 151, 4228–4238.

    CAS  PubMed  Google Scholar 

  101. Bourguignon, L. Y. W., Lokeshwar, V. B., He, J., Chen, X., Bourgignon, G. J. (1992) A CD44-like endothelial cell transmembrane glycoprotein (GP116) interacts with extracellular matrix and ankyrin. Mol. Cell. Biol. 12, 4461–4471.

    Google Scholar 

  102. Haugland, R. P. (1994) Spectra of fluorescent dyes used in flow cytometry. Methods Cell Biol. 42, 641–663.

    CAS  PubMed  Google Scholar 

  103. Milici, A. J. and Porter, G. A. (1991) Lectin and immunolabeling of microvascular endothelia. J. Electron Microsc. Tech. 19, 305–315.

    CAS  PubMed  Google Scholar 

  104. Tomczok, J., Saliwa-Tomczok, W., Klein, C. L., Bittinger, F., and Kirkpatrick, C. J. (1994) Application of immunogold labelling for light and electron microscopic localization of endothelial leukocyte adhesion molecule 1 (ELAM-1) on cultured human endothelial cells. Micron. 25, 257–266.

    CAS  PubMed  Google Scholar 

  105. Voyta, J. C., Via, D. P., Butterfield, C. E., and Zetter, B. R. (1984) Identtfication and isolation of endothelial cells based on their increased uptake of acetylated-low density lipoprotein. J. Cell Biol. 99, 2034–2040.

    CAS  PubMed  Google Scholar 

  106. Neumuller, J., Menzel, J., Neumark, T., Ferencz, G., and Tohidast-Akrad, M. (1992) Characterization of endothelial cell cultures derived from human synovial tissue, in Electron Microscopy, Vol. 3. Biological Sciences (Megías-Megías, L., Rodríguez-García, M. I., Ríos, A., and Arias, J. M., eds.), EUREM, Granada Secretariado de Publicaciones de la Universidad de Granada, pp. 531–532.

    Google Scholar 

  107. Cramer, C. T. (1989) An ultrathin sectioning alignment tool with application to cell monolayers. J. Electron Microsc. Tech. 11, 172–173.

    CAS  PubMed  Google Scholar 

  108. Arnold, J. R. and Boor, P. J. (1986) Improved transmission electron microscopy (TEM) of cultured cells through a “floating sheet” method. J. Ultrastruct. Mol. Struct. Res. 94, 30–36.

    CAS  PubMed  Google Scholar 

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  1. Ludwig Boltzmann Institute of Rheumatology and Balneology, Vienna, Austria

    Josef Neumüller & Johannes Menzel

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  1. Josef Neumüller
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  1. Albert Einstein College of Medicine, Bronx, NY

    Jeffrey W. Pollard

  2. University of Hertfordshire, Hatfield, Herts, UK

    John M. Walker

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Neumüller, J., Menzel, J. (1997). Adhesion Molecules on Isolated and Cultured Microvascular Endothelial Cells Demonstrated by Immunofluorescence and Immune Electron Microscopy. In: Pollard, J.W., Walker, J.M. (eds) Basic Cell Culture Protocols. Methods in Molecular Biology™, vol 75. Humana Press, Totowa, NJ. https://doi.org/10.1385/0-89603-441-0:399

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  • DOI: https://doi.org/10.1385/0-89603-441-0:399

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