Long-Term Co-Cultures of Adult Human Hepatocytes with Rat Liver Epithelial Cells: Modulation of Albumin Secretion and Accumulation of Extracellular Material

Authors

  • Bruno Clement,

    1. Unité de Recherches Hépatologiques U-49 de l'INSERM and Clinique Chirurgicale B, Hǒpital Pontchaillou, 35011 Rennes Cedex, France
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  • Christiane Guguen-Guillouzo,

    Corresponding author
    1. Unité de Recherches Hépatologiques U-49 de l'INSERM and Clinique Chirurgicale B, Hǒpital Pontchaillou, 35011 Rennes Cedex, France
    • Christiane Guguen-Guillouzo, Ph.D., Unité de Recherches Hépatologiques U-49 de l'INSERM, Hǒpital Pontchaillou, 35011 Rennes Cédex, France.
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  • Jean-Pierre Campion,

    1. Unité de Recherches Hépatologiques U-49 de l'INSERM and Clinique Chirurgicale B, Hǒpital Pontchaillou, 35011 Rennes Cedex, France
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  • Denise Glaise,

    1. Unité de Recherches Hépatologiques U-49 de l'INSERM and Clinique Chirurgicale B, Hǒpital Pontchaillou, 35011 Rennes Cedex, France
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  • Michel Bourel,

    1. Unité de Recherches Hépatologiques U-49 de l'INSERM and Clinique Chirurgicale B, Hǒpital Pontchaillou, 35011 Rennes Cedex, France
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  • Andre Guillouzo

    1. Unité de Recherches Hépatologiques U-49 de l'INSERM and Clinique Chirurgicale B, Hǒpital Pontchaillou, 35011 Rennes Cedex, France
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Abstract

High yields of viable human hepatocytes were obtained by enzymatic perfusion of the left hepatic lobe of kidney donors and cultured alone or with an epithelial cell line derived from rat liver. In conventional cultures, human hepatocytes did not survive more than 2 to 3 weeks and by Day 8 decreased their ability to secrete albumin. When co-cultured, they survived for more than 2 months and secreted high levels of albumin even in a serum-free medium. This long-term survival appeared to correlate with production of an extracellular material which is rich in Type III collagen. In vitro phenotypic alterations of parenchymal cells were reversed by addition of rat liver cells and were characterized by recovery of cuboidal morphology, increased albumin secretion and a shift from Type I to Type III collagen deposition. Rat liver epithelial cells could not be replaced by nonhepatic epithelial cells. These observations suggest that when adult human hepatocytes are maintained in a culture which closely resembles their in vivo environment, they are capable of continuing to actively express specific cell functions.

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