Skip to main content

Some NLM-NCBI services and products are experiencing heavy traffic, which may affect performance and availability. We apologize for the inconvenience and appreciate your patience. For assistance, please contact our Help Desk at info@ncbi.nlm.nih.gov.

The Journal of Clinical Investigation logoLink to The Journal of Clinical Investigation
. 1992 Oct;90(4):1284–1289. doi: 10.1172/JCI115992

Human intrahepatic biliary epithelial cells proliferate in vitro in response to human hepatocyte growth factor.

R Joplin 1, T Hishida 1, H Tsubouchi 1, Y Daikuhara 1, R Ayres 1, J M Neuberger 1, A J Strain 1
PMCID: PMC443171  PMID: 1401065

Abstract

In previous studies, intrahepatic human biliary epithelial cells (BEC) were isolated in high purity. However, these cells demonstrated only limited growth responses. Here we report that human BEC proliferate in response to human hepatocyte growth factor (hHGF), retain BEC-specific phenotype, and can be serially passaged. BEC showed dose-dependent growth in response to 0.01-100 ng/ml hHGF. The maximum S-phase labeling index reached 40% with half-maximal stimulation at 1 ng/ml. The response of cells from normal and primary biliary cirrhotic liver to hHGF was similar. Cultures were immunostained with specific antibodies and then processed for [3H]thymidine autoradiography. Proliferating cells expressed BEC-specific markers (HEA125 and CK-19), but were negative for desmin and factor VIII-related antigen. Occasional vimentin-positive cells were observed, but these were nonproliferative. In conclusion, cells responding to hHGF were clearly BEC in origin. The observation that HGF is mitogenic for BEC as well as hepatocytes has important implications. First, greater yields of intrahepatic BEC are available for subsequent studies of the pathogenesis and etiology of diseases of the biliary epithelium. Secondly, some means of regulating the cellular response to HGF in vivo must operate, in that HGF levels rise early after partial hepatectomy and yet BEC proliferate 24 h later than hepatocytes.

Full text

PDF
1284

Images in this article

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Ayres R. C., Shaw J., Mills C. O., Coleman R., Neuberger J. M. A 51Cr release cytotoxicity assay for use with human intrahepatic biliary epithelial cells. J Immunol Methods. 1991 Jul 26;141(1):117–122. doi: 10.1016/0022-1759(91)90217-4. [DOI] [PubMed] [Google Scholar]
  2. Belzer F. O., Southard J. H. Principles of solid-organ preservation by cold storage. Transplantation. 1988 Apr;45(4):673–676. doi: 10.1097/00007890-198804000-00001. [DOI] [PubMed] [Google Scholar]
  3. Ben-Ze'ev A. Differential control of cytokeratins and vimentin synthesis by cell-cell contact and cell spreading in cultured epithelial cells. J Cell Biol. 1984 Oct;99(4 Pt 1):1424–1433. doi: 10.1083/jcb.99.4.1424. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Burgess W. H., Maciag T. The heparin-binding (fibroblast) growth factor family of proteins. Annu Rev Biochem. 1989;58:575–606. doi: 10.1146/annurev.bi.58.070189.003043. [DOI] [PubMed] [Google Scholar]
  5. Fausto N., Mead J. E. Regulation of liver growth: protooncogenes and transforming growth factors. Lab Invest. 1989 Jan;60(1):4–13. [PubMed] [Google Scholar]
  6. Gohda E., Tsubouchi H., Nakayama H., Hirono S., Sakiyama O., Takahashi K., Miyazaki H., Hashimoto S., Daikuhara Y. Purification and partial characterization of hepatocyte growth factor from plasma of a patient with fulminant hepatic failure. J Clin Invest. 1988 Feb;81(2):414–419. doi: 10.1172/JCI113334. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Gröne H. J., Weber K., Gröne E., Helmchen U., Osborn M. Coexpression of keratin and vimentin in damaged and regenerating tubular epithelia of the kidney. Am J Pathol. 1987 Oct;129(1):1–8. [PMC free article] [PubMed] [Google Scholar]
  8. Igawa T., Kanda S., Kanetake H., Saitoh Y., Ichihara A., Tomita Y., Nakamura T. Hepatocyte growth factor is a potent mitogen for cultured rabbit renal tubular epithelial cells. Biochem Biophys Res Commun. 1991 Jan 31;174(2):831–838. doi: 10.1016/0006-291x(91)91493-v. [DOI] [PubMed] [Google Scholar]
  9. Ismail T., Howl J., Wheatley M., McMaster P., Neuberger J. M., Strain A. J. Growth of normal human hepatocytes in primary culture: effect of hormones and growth factors on DNA synthesis. Hepatology. 1991 Dec;14(6):1076–1082. [PubMed] [Google Scholar]
  10. Joplin R., Lindsay J. G., Johnson G. D., Strain A., Neuberger J. Membrane dihydrolipoamide acetyltransferase (E2) on human biliary epithelial cells in primary biliary cirrhosis. Lancet. 1992 Jan 11;339(8785):93–94. doi: 10.1016/0140-6736(92)91001-o. [DOI] [PubMed] [Google Scholar]
  11. Joplin R., Strain A. J., Neuberger J. M. Biliary epithelial cells from the liver of patients with primary biliary cirrhosis: isolation, characterization, and short-term culture. J Pathol. 1990 Nov;162(3):255–260. doi: 10.1002/path.1711620312. [DOI] [PubMed] [Google Scholar]
  12. Joplin R., Strain A. J., Neuberger J. M. Immuno-isolation and culture of biliary epithelial cells from normal human liver. In Vitro Cell Dev Biol. 1989 Dec;25(12):1189–1192. doi: 10.1007/BF02621273. [DOI] [PubMed] [Google Scholar]
  13. Kan M., Zhang G. H., Zarnegar R., Michalopoulos G., Myoken Y., McKeehan W. L., Stevens J. I. Hepatocyte growth factor/hepatopoietin A stimulates the growth of rat kidney proximal tubule epithelial cells (RPTE), rat nonparenchymal liver cells, human melanoma cells, mouse keratinocytes and stimulates anchorage-independent growth of SV-40 transformed RPTE. Biochem Biophys Res Commun. 1991 Jan 15;174(1):331–337. doi: 10.1016/0006-291x(91)90524-b. [DOI] [PubMed] [Google Scholar]
  14. Kasper M., Karsten U. Coexpression of cytokeratin and vimentin in Rathke's cysts of the human pituitary gland. Cell Tissue Res. 1988 Aug;253(2):419–424. doi: 10.1007/BF00222299. [DOI] [PubMed] [Google Scholar]
  15. Kinoshita T., Tashiro K., Nakamura T. Marked increase of HGF mRNA in non-parenchymal liver cells of rats treated with hepatotoxins. Biochem Biophys Res Commun. 1989 Dec 29;165(3):1229–1234. doi: 10.1016/0006-291x(89)92733-2. [DOI] [PubMed] [Google Scholar]
  16. Lindroos P. M., Zarnegar R., Michalopoulos G. K. Hepatocyte growth factor (hepatopoietin A) rapidly increases in plasma before DNA synthesis and liver regeneration stimulated by partial hepatectomy and carbon tetrachloride administration. Hepatology. 1991 Apr;13(4):743–750. [PubMed] [Google Scholar]
  17. Milani S., Herbst H., Schuppan D., Niedobitek G., Kim K. Y., Stein H. Vimentin expression of newly formed rat bile duct epithelial cells in secondary biliary fibrosis. Virchows Arch A Pathol Anat Histopathol. 1989;415(3):237–242. doi: 10.1007/BF00724910. [DOI] [PubMed] [Google Scholar]
  18. Momburg F., Moldenhauer G., Hämmerling G. J., Möller P. Immunohistochemical study of the expression of a Mr 34,000 human epithelium-specific surface glycoprotein in normal and malignant tissues. Cancer Res. 1987 Jun 1;47(11):2883–2891. [PubMed] [Google Scholar]
  19. Nakamura T., Nawa K., Ichihara A. Partial purification and characterization of hepatocyte growth factor from serum of hepatectomized rats. Biochem Biophys Res Commun. 1984 Aug 16;122(3):1450–1459. doi: 10.1016/0006-291x(84)91253-1. [DOI] [PubMed] [Google Scholar]
  20. Prat M., Narsimhan R. P., Crepaldi T., Nicotra M. R., Natali P. G., Comoglio P. M. The receptor encoded by the human c-MET oncogene is expressed in hepatocytes, epithelial cells and solid tumors. Int J Cancer. 1991 Sep 30;49(3):323–328. doi: 10.1002/ijc.2910490302. [DOI] [PubMed] [Google Scholar]
  21. Rheinwald J. G., Green H. Serial cultivation of strains of human epidermal keratinocytes: the formation of keratinizing colonies from single cells. Cell. 1975 Nov;6(3):331–343. doi: 10.1016/s0092-8674(75)80001-8. [DOI] [PubMed] [Google Scholar]
  22. Rosen E. M., Jaken S., Carley W., Luckett P. M., Setter E., Bhargava M., Goldberg I. D. Regulation of motility in bovine brain endothelial cells. J Cell Physiol. 1991 Feb;146(2):325–335. doi: 10.1002/jcp.1041460218. [DOI] [PubMed] [Google Scholar]
  23. Rubin J. S., Chan A. M., Bottaro D. P., Burgess W. H., Taylor W. G., Cech A. C., Hirschfield D. W., Wong J., Miki T., Finch P. W. A broad-spectrum human lung fibroblast-derived mitogen is a variant of hepatocyte growth factor. Proc Natl Acad Sci U S A. 1991 Jan 15;88(2):415–419. doi: 10.1073/pnas.88.2.415. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Rubin J. S., Osada H., Finch P. W., Taylor W. G., Rudikoff S., Aaronson S. A. Purification and characterization of a newly identified growth factor specific for epithelial cells. Proc Natl Acad Sci U S A. 1989 Feb;86(3):802–806. doi: 10.1073/pnas.86.3.802. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Saidman S. L., Duquesnoy R. J., Zeevi A., Fung J. J., Starzl T. E., Demetris A. J. Recognition of major histocompatibility complex antigens on cultured human biliary epithelial cells by alloreactive lymphocytes. Hepatology. 1991 Feb;13(2):239–246. [PMC free article] [PubMed] [Google Scholar]
  26. Sirica A. E., Mathis G. A., Sano N., Elmore L. W. Isolation, culture, and transplantation of intrahepatic biliary epithelial cells and oval cells. Pathobiology. 1990;58(1):44–64. doi: 10.1159/000163564. [DOI] [PubMed] [Google Scholar]
  27. Stanley M. A., Parkinson E. K. Growth requirements of human cervical epithelial cells in culture. Int J Cancer. 1979 Oct 15;24(4):407–414. doi: 10.1002/ijc.2910240406. [DOI] [PubMed] [Google Scholar]
  28. Strain A. J., Ismail T., Tsubouchi H., Arakaki N., Hishida T., Kitamura N., Daikuhara Y., McMaster P. Native and recombinant human hepatocyte growth factors are highly potent promoters of DNA synthesis in both human and rat hepatocytes. J Clin Invest. 1991 May;87(5):1853–1857. doi: 10.1172/JCI115207. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Strain A. J., McGowan J. A., Bucher N. L. Stimulation of DNA synthesis in primary cultures of adult rat hepatocytes by rat platelet-associated substance(s). In Vitro. 1982 Feb;18(2):108–116. doi: 10.1007/BF02796402. [DOI] [PubMed] [Google Scholar]
  30. Tashiro K., Hagiya M., Nishizawa T., Seki T., Shimonishi M., Shimizu S., Nakamura T. Deduced primary structure of rat hepatocyte growth factor and expression of the mRNA in rat tissues. Proc Natl Acad Sci U S A. 1990 Apr;87(8):3200–3204. doi: 10.1073/pnas.87.8.3200. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Van Eyken P., Sciot R., Desmet V. Intrahepatic bile duct development in the rat: a cytokeratin-immunohistochemical study. Lab Invest. 1988 Jul;59(1):52–59. [PubMed] [Google Scholar]
  32. Weidner K. M., Arakaki N., Hartmann G., Vandekerckhove J., Weingart S., Rieder H., Fonatsch C., Tsubouchi H., Hishida T., Daikuhara Y. Evidence for the identity of human scatter factor and human hepatocyte growth factor. Proc Natl Acad Sci U S A. 1991 Aug 15;88(16):7001–7005. doi: 10.1073/pnas.88.16.7001. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Wolf H. K., Zarnegar R., Michalopoulos G. K. Localization of hepatocyte growth factor in human and rat tissues: an immunohistochemical study. Hepatology. 1991 Sep;14(3):488–494. [PubMed] [Google Scholar]
  34. Yoshitomi S., Miyazaki K., Nakayama F. Demonstration and maintenance of mucus secretion in cultured human gallbladder epithelial cells. In Vitro Cell Dev Biol. 1987 Aug;23(8):559–566. doi: 10.1007/BF02620973. [DOI] [PubMed] [Google Scholar]
  35. Zarnegar R., DeFrances M. C., Kost D. P., Lindroos P., Michalopoulos G. K. Expression of hepatocyte growth factor mRNA in regenerating rat liver after partial hepatectomy. Biochem Biophys Res Commun. 1991 May 31;177(1):559–565. doi: 10.1016/0006-291x(91)92020-k. [DOI] [PubMed] [Google Scholar]
  36. Zarnegar R., Michalopoulos G. Purification and biological characterization of human hepatopoietin A, a polypeptide growth factor for hepatocytes. Cancer Res. 1989 Jun 15;49(12):3314–3320. [PubMed] [Google Scholar]
  37. van Eyken P., Sciot R., van Damme B., de Wolf-Peeters C., Desmet V. J. Keratin immunohistochemistry in normal human liver. Cytokeratin pattern of hepatocytes, bile ducts and acinar gradient. Virchows Arch A Pathol Anat Histopathol. 1987;412(1):63–72. doi: 10.1007/BF00750732. [DOI] [PubMed] [Google Scholar]

Articles from Journal of Clinical Investigation are provided here courtesy of American Society for Clinical Investigation

RESOURCES