Characterization of human hepatocyte lines derived from normal liver tissue

Authors

  • Eve A. Roberts M.D.,

    Corresponding author
    1. Division of Gastroenterology, Toronto, Ontario, Canada
    2. Division of Clinical Pharmacology and Toxicology, Toronto, Ontario, Canada
    • Division of Gastroenterology, Room 8431, Gerrard Wing, The Hospital for Sick Children, 555 University Avenue, Toronto, Ontario M5G 1X8, Canada
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  • Michelle Letarte,

    1. Division of Immunology and Cancer Research, The Hospital for Sick Children Research Institute, Toronto, Ontario, Canada
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  • Jeremy Squire,

    1. Department of Pathology, The Hospital for Sick Children, and the University of Toronto, Toronto, Ontario, Canada
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  • Suyun Yang

    1. Division of Gastroenterology, Toronto, Ontario, Canada
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Abstract

Four separate continuous lines of human hepatocytes (HH01, HH02, HH09, HH25) were developed from normal liver tissue by subjecting cocultures of human hepatocytes with rat liver epithelial cells in a highly enriched medium to frequent subculturing. The addition of conditioned medium from either the human hepatoma line Hep G2 or one of these stable human hepatocyte lines (HH09) appeared to facilitate establishment of line HH25. These human hepatocyte lines have been in continuous culture for 2 to 5 yr and, consist of approximately 95% human cells by analysis of cell surface antigens. Cytogenetic analysis also confirmed the human origin of these cells and showed clonal origin with abnormal ploidy. Cells in these human hepatocyte lines retain morphological features of hepatocytes by both light and electron microscopy. They also retain glucose-6-phosphatase activity and secrete proteins characteristic of hepatocytes, such as albumin, α-fetoprotein and transferrin. After incubation with 13 μmol/L dihenz(a,h) anthracene for 24 hr, each line had detectable activity of aryl hydrocarbon hydroxylase, ethoxyresorufin O-deethylase and methoxyresorufin O-demethylase. Thus, these human hepatocyte lines retain important differentiated characteristics of hepatocytes. Derived from normal liver tissue, they appear to be immortalized. They provide a new model system for studying human hepatocellular drug metabolism. These lines may also be useful for studying the regulation of synthesis of albumin, α-fetoprotein and other proteins in human hepatocytes, determining the effects of cytokines and growth factors and designing systems to effect gene transfer into human hepatocytes for the purpose of gene therapy. (HEPATOLOGY 1994;19:1390–1399.)

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