Skip to main content
SpringerLink
Log in

Monoclonal antibodies directed against rat liver epithelial cell lines selectively recognize bile duct epithelium in livers of adult rats

  • Published:
Cell Biology and Toxicology Aims and scope Submit manuscript

Monoclonal antibodies directed against antigens on rat liver epithelial cell lines were prepared. Three antibodies, 4C3, 19C6, and 3C2, recognized surface antigens present (although in different quantities) on eight epithelial cell lines tested, irrespective of whether they were normal or transformed. For MAb 3C2, the primary antigen common to all but one cell line showed a Mr of 135 kD. In paraffin sections of liver tissue, two antibodies, 40 and 19C6, reacted exclusively with bile duct epithelium, whereas the MAb 3C2 additionally reacted with sinusoidal endothelium and the endothelium of the portal venules. In sections of livers from rats exposed to diethylnitrosamine, the MAb 19C6 selectively stained bile duct-like structures in cholangiomas, while other preneoplastic and neoplastic lesions were not stained. These results demonstrate that the monoclonal antibodies obtained may prove useful for investigating cell lineages related to propagable liver epithelial cell lines and suggest that these cells may be derived from terminal bile ductular cells.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

Abbreviations

ABTS2:

2,2′azinobis(3-ethylbenzthiazolinesulfonic) acid

ARL:

adult rat liver

DEN:

diethylnitrosamine

FCS:

fetal calf serum

MAb:

monoclonal antibody

PAP:

peroxidase antiperoxidase

References

  • ADEN, D.P., FOGEL, A., PLOTKIN, S., DAMJANOV, I. and KNOWLES, B.B. (1979). Controlled synthesis of HBsAg in a differentiated human liver carcinoma-derived cell line. Nature 282:615–616.

    Google Scholar 

  • BOREK, C. (1972). Neoplastic transformation in vitro of a clone of adult rat liver epithelial cells into differentiated hepatoma-like cells under conditions of nutritional stress. Proc. Natl. Acad. Sci. USA 69:956–959.

    Google Scholar 

  • BURGER, H.-J. (1987). Heterogenität des Glutaminstoffwechsels in der Rattenleber. Thesis, Tübingen.

  • CHESSEBEUF, M., EXILIE, M.F., PIRARD, G., CHATELAIN, P. and PADIEU, P. (1981). Effect of serum concentration on the chromosome number of a rat liver epithelial cell line. Cell Biol. Int. Rep. 5:399–408.

    Google Scholar 

  • DOUILLARD, J.Y. and HOFFMAN, T. (1983). Enzyme-linked immunosorbent assay for screening monoclonal antibody production using enzyme-labeled second antibody. Meth. Enzymol. 92:168.

    Google Scholar 

  • FURUKAWA, K., SHIMADA, T., ENGLAND, P., MOCHIZUKI, Y. and WILLIAMS, G.M. (1987). Enrichment and characterization of clonogenic epithelial cells from adult rat liver and initiation of epithelial cell strains. In Vitro Cell. Devel. Biol. 23:339–348.

    Google Scholar 

  • GEBHARDT, R. and BURGER, H.-J. (1987). Selective loss of hormonal induction of glutamate transport in [rimary cultures of hepatocytes from rats treated with CCI4. J. Hepatol. 4:381–389.

    Google Scholar 

  • GEBHARDT, R. and JUNG, W. (1982). Biliary secretion of sodium fluorescein in primary monolayer cultures of rat hepatocytes and its stimulation by nicotinamide. J. Cell Sci. 56:233–244.

    Google Scholar 

  • GEBHARDT, R. and MECKE, D. (1983). Heterogeneous distribution of glutamine synthetase among rat liver parenchymal cells in situ and in primary culture.EMBO J. 2:567–570.

    Google Scholar 

  • GEBHARDT, R. and WILLIAMS, G.M. (1986). Amino acid transport in established adult rat liver epithelial cell lines. Cell Biol. Toxicol. 2:9–20.

    Google Scholar 

  • GEBHARDT, R. and WILLIAMS, G.M. (1988). A new aspect of liver carcinogenesis: selective induction of glutamine-synthetase-positive liver carcinomas by carcinogens which affect hepatocytes expressing this phenotype. In Advances in Ammonia Metabolism and Hepatic Encephalopathy (P.B. Soeters, J.H.P. Wilson, A.J. Meijer, E. Holm, eds.) pp. 581–586. Elsevier Science Publishers, Amsterdam.

    Google Scholar 

  • GERSCHENSON, L.E., ANDERSSON, M., MOLSON, J. and OKIGAKI, T. (1970). Tyrosine aminotransferase induction by dexamethasone in a new rat liver cell line. Science 170:859–861.

    Google Scholar 

  • GERSCHENSON, L.E. (1975). Hormonal effects on two rat liver cell lines cultured in chemically defined medium. In: Gene Expression and Carcinogenesis in Cultured Liver (L.E. Gerschenson, E.B. Thompson, eds.) pp. 220–226. Academic Press, New York.

    Google Scholar 

  • GRISHAM, J.W., THAL, S.B. and NAGEL, A. (1975). Cellular derivation of continuously cultured epithelial cells from normal rat liver. In: Gene Expression and Carcinogenesis in Cultured Liver (L.E. Gerschenson, E.B. Thompson, eds.) pp. 1–23. Academic Press, New York.

    Google Scholar 

  • GRISHAM, J.W. (1980). Cell types in long-term propagable cultures of rat liver. Ann. NY Acad. Sci. 349:128–137.

    Google Scholar 

  • HERRING, A.S., RAYCHAUDHURI, R. and KELLEY, S.P. (1983). Repeated establishment of diploid epithelial cell cultures from normal and partially hepatectomized rats. In Vitro 19:576–588.

    Google Scholar 

  • HIXON, D.C. and ALLISON, J.P. (1985). Monoclonal antibodies recognizing oval cells induced in the liver of rats by n-2-fluorenylacetamide or ethionine in a choline-deficient diet. Cancer Res. 45:3750–3759.

    Google Scholar 

  • INAOKA, Y. (1967). Significance of the so-called oval cell proliferation during azo-dye hepatocarcinogenesis. Gann 58:355–366.

    Google Scholar 

  • ISMAIL-BEIGI, F., HABER, R.S. and LOEB, J.H. (1986). Stimulation of active NA+and K+ transport by thyroid hormone in a rat liver cell line: role of enhanced Na+ entry. Endocrinology 119:2527–2536.

    Google Scholar 

  • IYPE, P.T. (1971). Cultures from adult rat liver cells. I. Establishment of monolayer cell cultures from normal liver. J. Cell. Physiol. 78:281–288.

    Google Scholar 

  • IYPE, P.T., BALDWIN, R.W. and CLAVIS, D. (1972). Cultures from adult rat liver cells. II. Demonstration of organ specific cell surface antigens on cultured cells from normal liver. Br. J. Cancer 26:6–9.

    Google Scholar 

  • JOHANSSON, S., GUSTAFSON, S. and PERTOFT, H. (1987). Identification of a fibronectin receptor specific for rat liver endothelial cells. Exp. Cell Res. 172:425–431.

    Google Scholar 

  • JUNG, W., GEBHARDT, R. and MECKE, D. (1982). Alterations in activity and ultrastructural localization of several phosphatases on the surface of adult rat hepatocytes in primary monolayer culture. Eur. J. Cell Biol. 27:230–241.

    Google Scholar 

  • KÖHLER, G. and MILSTEIN, C. (1975). Continuous cultures of fused cells secreting antibody of predefined specificity. Nature 256:495–497.

    Google Scholar 

  • KUHLMANN, W.D. (1978). Localization of alpha-fetoprotein and DNA-synthesis in liver cell populations during experimental hepatocarcinogenesis in rats. Int. J. Cancer 21:368–380.

    Google Scholar 

  • LAEMMLI, U.K. (1970). Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680–685.

    Google Scholar 

  • LANDSDORP, P.M. (1984). Stepwise amplified immunoperoxidase staining. J. Histochem. Cytochem. 32:172–178.

    Google Scholar 

  • LOMBARDI, B. (1982). On the nature, properties and significance of oval cells, biological mechanisms and environmental factors in pathology. In: Recent Trends in Chemical Carcinogenesis (P. Pani, F. Feo and A. Culumbano, eds.) Vol. 1, pp. 37–56. E.S.A., Cagliari.

    Google Scholar 

  • MAYER, D., and SCHAFER, B. (1982). Biochemical and morphological characterization of glycogen-storing epithelial liver cell lines. Exp. Cell Res. 138:1–14.

    Google Scholar 

  • MARCEAU, N., GERMAIN, L., GOYETTE, R., NOEL, M. and GOURDEAU, H. (1986). Cell of origin of distinct rat liver epithelial cells, as typed by cytokeratin and surface component selective expression. Biochem. Cell Biol. 64:788–802.

    Google Scholar 

  • NEUPERT, G., LANGBEIN, L. and KARSTEN, U. (1987). Characterization of established epitheloid cell lines derived from rat liver: expression of cytokeratin filaments. Exp Pathol. 31:161–167.

    Google Scholar 

  • ODASHIMA, S., STURGESS, J.M. and ROTHSTEIN, A. (1976). Studies with mature rat hepatocytes in monolayer culture. I. Propagation of isolated hepatocytes. Morphological and biochemical characteristics in culture. Cell Tissue Res. 169:167–178.

    Google Scholar 

  • OGAWA, H., MINASE, T. and ONOE, T. (1974). Demonstration of glucose-6-phophatase activity in the oval cells of rat liver and the significance of the oval cells in azo dye carcinogenesis. Cancer Res. 34:3379–3386.

    Google Scholar 

  • SAN, R.H.C., LASPIA, M.F., SOIEFER, A.I., MASLANSKY, C.J., RICE, J.M. and WILLIAMS, G.M. (1979). A survey of growth in soft agar and cell surface properties as markers for transformation in adult rat liver epithelial-like cell cultures. Cancer Res. 39:1026–1034.

    Google Scholar 

  • SAN, R.H.C., SHIMADA, T., MASLANSKY, C.J., KREISER, D.M. LASPIA, M.F., RICE, J.M. and WILLIAMS, G.M. (1979). Growth characteristics and enzyme activities in a survey of transformation markers in adult rat liver epithelial-like cell cultures. Cancer Res. 39:4441–4448.

    Google Scholar 

  • SELL, S. and LEFFERT, H.L. (1982). An evaluation of cellular lineages in the pathogenesis of experimental hepatocellular carcinoma. Hepatology 2:77–68.

    Google Scholar 

  • SHIMADA, T., FURUKAWA, K., KREISER, D.M., CAWEIN, A. and WILLIAMS, G.M. (1983). Induction of transformation by six classes of chemical carcinogens in adult rat liver epithelial cells. Cancer Res. 43:5087–5092.

    Google Scholar 

  • STERNBERGER, L.A. (1970). The unlabeled antibody enzyme method of immunohistochemistry. J. Histochem. Cytochem. 18:325–333.

    Google Scholar 

  • TONG, C. and WILLIAMS, G.M. (1978). Induction of purine analog-resistant mutants in adult rat liver epithelial lines by metabolic activation-dependent and -independent carcinogens. Mutat. Res. 58:339–352.

    Google Scholar 

  • TOWBIN, H., STAEHLIN, T. and GORDON, J. (1979). Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc. Natl. Acad. Sci. USA 76:4350–4354.

    Google Scholar 

  • TSAO, M.S., SMITH, J.D., NELSON, K.G. and GRISHAM, J.W. (1984). A diploid epithelial cell line from normal adult rat liver with phenotypic properties of “oval” cells. Exp. Cell Res. 154:38–52.

    Google Scholar 

  • TSAO, M.S., GRISHAM, J.W., NELSON, K.G. and SMITH, J.D. (1985a). Phenotypic and karyotypic alterations induced in cultured rat hepatic epithelial cells that express the “oval” cell phenotype by exposure to N-methyl-N′-nitro-N-nitrosoguanidine. Am. J. Pathol. 118:306–315.

    Google Scholar 

  • TSAO, M.S., GRISHAM, J.W. and NELSON, K.G. (1985b). Clonal analysis of tumorigenicity and paratumorigenic phenotypes in rat liver epithelial cells chemically transformed in vitro. Cancer Res. 45:5139–5144.

    Google Scholar 

  • WEINSTEIN, I.B., ORENSTEIN, J.M. and GEBERT, R. (1975). Growth and structural properties of epithelial cell cultures established from normal rat liver and chemically induced hepatomas. Cancer Res. 35:253–263.

    Google Scholar 

  • WILLIAMS, G.M. (1975). The study of chemical carcinogenesis using cultured rat liver cells. In Gene Expression and Carcinogenesis in Cultured Liver: (L.E. Gerschenson, E.B. Thompson, eds.) pp. 480–487. Academic Press, New York.

    Google Scholar 

  • WILLIAMS, G.M. (1976a). Primary and long term culture of adult rat liver epithelial cells. Methods Cell Biol. 14:357–364.

    Google Scholar 

  • WILLIAMS, G.M. (1976b). The use of liver epithelial cultures for the study of chemical carcinogenesis. Am. J. Pathol. 85:739–753.

    Google Scholar 

  • WILLIAMS, G.M. (1981). Mammalian culture systems for the study of genetic effects of N-substituted aryl compounds. Natl. Cancer Inst. Monogr. 58:237–242.

    Google Scholar 

  • WILLIAMS, G.M. and GUNN J.M. (1974). Long-term cell culture of adult rat liver epithelial cells. Exp. Cell Res. 89:139–142.

    Google Scholar 

  • WILLIAMS, G.M., WEISBURGER, E.K. and WEISBURGER, J.H. (1971). Isolation and long-term culture of epithelial-like cells from rat liver. Exp. Cell Res. 69:106–112.

    Google Scholar 

  • WILLIAMS, G.M., STROMBERG, K. and KROES, R. (1973). Cytochemical and ultrastructural alterations associated with confluent growth of epithelial-like cells from rat liver. Lab. Investigations 29:293–303.

    Google Scholar 

  • WILLIAMS, G.M., ELLIOT, J.M. and WEISBURGER, J.H. (1973). Carcinoma after malignant conversion in vitro of epithelial-like cells from rat liver following exposure to chemical carcinogens. Cancer Res. 33:606–612.

    Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Physiologisch-chemisches Institut der Universität, Tubingen, Federal Republic of Germany

    Rolf Gebhardt & Ingeborg Schäfer-Degenhart

Authors
  1. Rolf Gebhardt
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Ingeborg Schäfer-Degenhart
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Gebhardt, R., Schäfer-Degenhart, I. Monoclonal antibodies directed against rat liver epithelial cell lines selectively recognize bile duct epithelium in livers of adult rats. Cell Biol Toxicol 4, 379–392 (1988). https://doi.org/10.1007/BF00117767

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00117767

Key words

Search

Navigation