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Membrane characteristics of adult rat liver parenchymal cells in primary monolayer culture

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Summary

Parenchymal cells, isolated from normal adult rat liver (3×107 cells/g liver) by collagenase perfusion and maintained in nondividing monolayer culture, were employed to investigate cell surface properties of hepatocytes. Membrane transport systems for asialoorosomucoid (A-OM) and methotrexate (MTX) were lost rapidly in culture, whereas induction of tyrosine aminotransferase and transport of α-aminoisobutyrate actually increased during the first 3 days. Alterations in the membrane transport systems for A-OM and MTX reflected more generalized modifications of cell surface components induced during primary culture. Thus, the binding of concanavalin A (Con A) and wheat germ agglutinin (WGA) to cultured hepatocytes increased approximately 2-fold between 24 and 96 hr, and the incorporation of radioactive mannose and glucosamine into trichloroacetic acid-insoluble proteins increased 13-fold and 4-fold, respectively.

Plasma membranes were isolated from cultured hepatocytes and the major structural proteins and glycoproteins were analyzed by SDS-polyacrylamide gel electrophoresis. Membrane instability between 24 and 96 hr of culture was characterized by time-dependent alterations in specific polypeptides and extensive changes in Con A- and WGA-binding glycoproteins. Although addition of a complex hormone supplement to the medium increased the number of viable cells and sustained A-OM and MTX transport systems for 24 hr, it had no influence on the altered membrane protein and glycoprotein profiles observed in its absence.

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  1. Division of Laboratories and Research, New York State Department of Health, 12201, Albany, New York

    A. L. Tarentino & J. Galivan

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  1. A. L. Tarentino
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  2. J. Galivan
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This work was supported by Grant AG 00207 from the National Institute of Aging.

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Tarentino, A.L., Galivan, J. Membrane characteristics of adult rat liver parenchymal cells in primary monolayer culture. In Vitro 16, 833–846 (1980). https://doi.org/10.1007/BF02619420

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