Sinusoidal Endothelial Cells from Normal Guinea Pig Liver: Isolation, Culture and Characterization

Authors

  • R. Gideon Shaw,

    1. Departments of Internal Medicine, Pathology and Pharmacology, The University of Texas Health Science Center at Dallas, Dallas, Texas 75235
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  • Alice R. Johnson,

    1. Departments of Internal Medicine, Pathology and Pharmacology, The University of Texas Health Science Center at Dallas, Dallas, Texas 75235
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  • Werner W. Schulz,

    1. Departments of Internal Medicine, Pathology and Pharmacology, The University of Texas Health Science Center at Dallas, Dallas, Texas 75235
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  • Rainer N. Zahlten,

    1. Departments of Internal Medicine, Pathology and Pharmacology, The University of Texas Health Science Center at Dallas, Dallas, Texas 75235
    Current affiliation:
    1. Medical Department, Division for Clinical Research, Hoechst Aktiengesellschaft, Frankfurt, West Germany.
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  • Burton Combes

    Corresponding author
    1. Departments of Internal Medicine, Pathology and Pharmacology, The University of Texas Health Science Center at Dallas, Dallas, Texas 75235
    • Burton Combes, M.D., Liver Unit, Department Internal Medicine, University of Texas Health Science Center at Dallas, Southwestern Medical School, 5323 Harry Hines Boulevard, Dallas, Texas 75235.
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  • Parts of this work were presented at the 84th Annual Meeting of the American Gastroenterological Association in Washington, D.C. on May 25, 1983, and appear in abstract form (Gastroenterology 1983; 84:1396).

Abstract

Guinea pig nonparenchymal hepatic cells were isolated by enzymatic digestion and subsequent separation on a 17.5% metrizamide gradient. Endothelial cell and Kupffer cell-enriched fractions were separated by centrifugal elutriation. Viability of both cell fractions was approximately 80%. Endothelial cells were cultured on a substratum of guinea pig liver collagen and 1% gelatin (1:1). Freshly isolated and cultured sinusoidal endothelial cells contained Factor VIII R:antigen, angiotensin I converting enzyme activity, and they synthesized prostaglandins characteristic of other endothelial cells. Sieve plates were identified in both freshly isolated and cultured cells. Fresh endothelial cells and Kupffer cells formed Fc receptor-mediated rosettes with IgG-opsonized sheep red blood cells, but cultured endothelial cells did not. Only Kupffer cells demonstrated Fc and C3 receptor-mediated phagocytosis. These methods for isolating and culturing sinusoidal endothelial cells should permit further functional assessment of endothelial cells and their interrelationship with other sinusoidal lining cells.

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