Summary
Although most vascular models use large vessel endothelial cells from human umbilical veins, there is marked heterogeneity among endothelial cells from different vascular beds and organs. More accurate modeling of endothelial involvement in liver diseases, including metastasis, may result from the use of human hepatic sinusoidal endothelial cells. Liver resection specimens were sectioned, then treated with a 1.2 U/ml dispase solution. The tissue slurry was mechanically disaggregated and separated by centrifugation on a Percoll density gradient. Cells were then cultured in an endothelial-specific media with growth factors. These techniques resulted in a homogeneous monolayer consistent with endothelial cells by light microscopy. An endothelial origin was further confirmed by the expression of Factor VIII, binding of Ulex lectin, and uptake of acetylated low density lipoprotein. Electron microscopy showed transcellular fenestrations consistent with a sinusoidal origin. These human hepatic sinusoidal endothelial cells were then studied for expression of the adhesion molecules CD31/PECAM, CD34, E-selectin, ICAM-1, L-selectin, LFA-3, P-selectin, and VCAM-1 plus the binding of wheat germ agglutinin lectin. The patterns of adhesion molecule expression and lectin binding by these cells are characteristic of hepatic sinusoidal endothelia. In this paper, we have described a method for isolation and culture of human cells with the morphologic and phenotypic characteristics of hepatic sinusoidal endothelia.
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Ades, E. J.; Candal, F. J.; Swerlick, R. A., et al. HMEC-1: establishment of an immortal human microvascular endothelial cell line. J. Invest. Dermatol. 99:683–690; 1992.
Apkarian, R. P. The Fine Structure of Fenestraeted Adrenocortical Capillaries Revealed by In-lens Field-emission Scanning Electron Microscopy and Scanning Transmission Electron Microscopy. Scanning 19:361–367; 1997.
Barbera-Guillem, E.; Alonso-Varona, A.; Vidal-Vanaclocha, F. Selective implantation of experimental hepatic metastasis in acinar zone one. Cancer Res. 49:4003–4010; 1989.
Barbera-Guillem, E.; Rocha, M.; Alvarez, A., et al. Differences in the lectin-binding patterns of the periportal and perivenous endothelial domains in the liver sinusoids. Hepatology 14:131–139; 1991.
Belloni, P. N.; Carney, D. H.; Nicholson, G. L. Organ-derived endothelial cells exhibit differential responsiveness to thrombin and other growth factors. Microvasc. Research 43:20–45; 1992.
Bowman, P. D.; Betz, A.; Ar, D., et al. Primary culture of capillary endothelium from rat brain. In Vitro Cell. Dev. Biol. 17:353; 1981.
Brouwer, A.; de Leeuw, A. W.; Praaning-van Dalen, D. P., et al. Isolation and culture of sinusoidal liver cells. In: W. E. Knook DL, ed. Sinusoidal liver cells. Amsterdam, Netherlands: Elsevier Biomedical Press; 1982:509–517.
Couvelard, A.; Scoazec, J. Y.; Feldmann, G. Expression of cell-cell and cell-matrix adhesion proteins by sinusoidal endothelial cells in the normal and cirrhotic human liver. Am. J. Pathol. 142:738–752; 1993.
Davison, P. M.; Bensch, K.; Karasek, M. A. Isolation and growth of endothelial cells from the microvessels of the newborn human foreskin in cell culture. J. Invest. Dermatol. 75:316; 1980.
Dorovini-Zis, K.; Prameya, R.; Bowman, P. D. Culture and characterization of microvascular endothelial cells derived from human brain. Lab. Invest. 63:425–436; 1991.
Folkman, J. C.; Hadenchild, C. C.; Zetter, B. R. Long term culture of capillary endothelial cells. Proc. Natl. Acad. Sci. USA 76:5217; 1979.
Gimbrone, M. A. Culture of vascular endothelium. Prog. Hemostasis Thromb. 3:1; 1976.
Goerdt, S.; Steckel, F.; Schulze-Osthoff, K., et al. Characterization and differential expression of an endothelial cell-specific surface antigen in continuous and sinusoidal endothelia, in skin vascular lesions and in vitro. Exp. Cell Biol. 57:185–192; 1989.
Goerdt, S.; Walsh, L. J.; Murphy, G. F., et al. Identification of a novel high molecular weight protein preferentially expressed by sinusoidal endothelial cells in normal human tissues. J. Cell Biol. 113:1425–1437; 1991.
Green, D. F.; Resnick, L.; Bourgoignie, J. J. HIV infects glomerular endothelial and mesangial but not epithelial cells in vitro. Kidney Int. 41:956–960; 1992.
Hormia, M.; Lehto, V. P.; Virtanen, I. Identification of UEA I binding surface glycoproteins of cultured endothelial cells. Cell Biol. Int. Rep. 7:467; 1983.
Jaffee, E. A. Endothelial cells and the biology of factor VIII. N. Engl. J. Med. 296:377; 1977.
Lafon, M. E.; Gendrault, J. L.; Royer, C., et al. Human endothelial cells isolated from the hepatic sinusoids and the umbilical vein display a different permissiveness for HIV1. Res. Virol. 144:99–104; 1993.
Lawley, T. J.; Kubota, Y. Cutaneous microvascular endothelial cells. In: Ryan, U. S., ed. Endothelial cells. Vol. III. Boca Raton, FL: CRC Press; 1992:229–240.
Rappaport, A. M. The microcirculatory hepatic unit. Microvasc. Res. 6:212–228; 1973.
Ryan, U. S.; Clements, E.; Habilston, D., et al. Isolation and culture of pulmonary artery endothelial cells. Tissue & Cell 10:535–554; 1978.
Scoazec, J.-Y.; Feldmann, G. In situ immunophenotyping study of endothelial cells of the human hepatic sinusoid: results and functional implications. Hepatology 14:789–797; 1991.
Scoazec, J.-Y.; Feldmann, G. The cell adhesion molecules of hepatic sinusoidal endothelial cells. J. Hepatol. 20:296–300; 1994.
Soda, R.; Tavassoli, M. Liver endothelium and not hepatocytes of Kupffer cells have transferrin receptors. Blood 63:270–276; 1984.
Steffan, A.-M.; Gendralt, J. L.; McCuskey, R. S., et al. Phagocytosis, an unrecognized property of murine endothelial liver cells. Hepatology 6:830–836; 1986.
Steffan, A.-M.; Lafon, M.-E.; Gendrault, J.-L., et al. Primary cultures of endothelial cells from the human liver sinusoid are permissive for human immunodeficiency virus type 1. Proc. Natl. Acad. Sci. 89:1528–1586; 1992.
Steffan, A.-M.; Lecerf, F.; Keller, F., et al. Isolement et culture de cellules endotheliliales de foies humain et murin. C. R. Acad. Sci (Series III) (Paris) 292:809–815; 1981.
Steinhoff, G.; Behrend, M.; Schrader, B. Intercellular immune adhesion molecules in human liver transplants: overview on expression patterns of leukocyte receptor and ligand molecules. Hepatology 18:440–453; 1993.
Steinhoff, G.; Behrend, M.; Schrader, B., et al. Expression patterns of leukocyte adhesion ligand molecules on human liver endothelia. Am. J. Pathol. 142:481–488; 1993.
Turner, R. R.; Beckstead, J. H.; Warnke, R. A., et al. Endothelial cell phenotypic diversity. Am. J. Clin. Pathol. 87:569–575; 1987.
Vidal-Vanaclocha, F.; Alonso-Varona, A.; Ayala, R., et al. Coincident implantation, growth and interaction sites within the liver of cancer and reactive hematopoietic cells. Int. J. Cancer 46:267–271; 1990.
Vidal-Vanaclocha, F.; Barbera-Guillem, E. Fenestration patterns in endothelial cells of rat liver sinusoids. J. Ultrastruct. Res. 90:115–123; 1985.
Vidal-Vanaclocha, F.; Rocha, M. A.; Asumendi, A., et al. Role of periportal and perivenous endothelial cells in hepatic homing of blood and metastatic cancer cells. Semin. Liver Dis. 13:60–71; 1993.
Volpes, R.; van den Oord, J. J.; Desmet, V. J. Vascular adhesion molecules in acute and chronic liver inflammation. Hepatology 15:269–275; 1992.
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Daneker, G.W., Lund, S.A., Caughman, S.W. et al. Culture and characterization of sinusoidal endothelial cells isolated from human liver. In Vitro Cell.Dev.Biol.-Animal 34, 370–377 (1998). https://doi.org/10.1007/s11626-998-0018-9
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DOI: https://doi.org/10.1007/s11626-998-0018-9