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A novel monoclonal antibody identified hepatic stem-like cells in rats

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Abstract

Both liver epithelial and oval cells are believed to be liver stem cells. We investigated the identification by producing monoclonal antibodies against liver epithelial cells. Monoclonal antibodies against hepatic stem-like cells (HSL cells) have been selected to follow the hepatic stem cells during hepatic regeneration and developmental changes in the liver. Monoclonal antibodies were induced by immunization of BALB/c mice with HSL cells established from the epithelial cells of the adult rat liver. The hybridomas were screened by indirect immunofluorescence staining of HSL cells. We produced a unique monoclonal antibody against HSL cells, MabH, which specifically recognizes liver epithelial cells. MabH did not react with liver parenchymal cells but did react with bile ductule cells under normal conditions in the adult liver. This antibody also reacted with oval cell lines and with the oval cells that appeared during liver regeneration. In addition, fetal liver cells showed immunoreactivity with MabH. Although the level of staining decreased after birth, some cells in the portal area remained highly reactive. These results suggested that liver epithelial cells, oval cells, and fetal liver cells possess a common cell marker of liver stem cells.

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References

  1. Tsao MS, Grisham JW, Nelson KG, Smith JD (1985) Phenotypic and karyotypic changes 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

    PubMed  CAS  Google Scholar 

  2. Maggie HW, David AG (2006) Hepatic stem cells: in search of. Stem Cells 24:1833–1840

    Article  Google Scholar 

  3. Santoni-Rugiu E, Jelnes P, Thorgeirsson SS, Bisgaard HC (2005) Progenitor cells in liver regeneration: molecular responses controlling their activation and expansion. APMIS 113:876–902

    Article  PubMed  Google Scholar 

  4. Paku S, Schnur J, Nagy P, Thorgeirsson SS (2001) Origin and structural evolution of the early proliferating oval cells in rat liver. Am J Pathol 158:1313–1323

    Article  PubMed  CAS  Google Scholar 

  5. Lazaro CA, Rhim JA, Yamada Y, Fausto N (1998) Generation of hepatocytes from oval cell precursors in culture. Cancer Res 58:5514–5522

    PubMed  CAS  Google Scholar 

  6. Yang L, Li S, Hatch H, Ahrens K, Cornelius JG, Petersen BE, Peck AB (2002) In vitro trans-differentiation of adult hepatic stem cells into pancreatic endocrine hormone-producing cells. Proc Natl Acad Sci U S A 99:8078–8083

    Article  PubMed  CAS  Google Scholar 

  7. Yasui O, Miura N, Terada K, Kawarada Y, Koyama K, Sugiyama T (1997) Isolation of oval cells from Long-Evans-Cinnamon rats and their transformation into hepatocytes in vivo in the rat liver. Hepatology 25:329–334

    PubMed  CAS  Google Scholar 

  8. Petersen BE, Bowen WC, Patrene KD, Mars WM, Sullivan AK, Murase N, Boggs SS, Greenberger JS, Goff JP (1999) Bone marrow as a potential source of hepatic oval cells. Science 284:1168–1170

    Article  PubMed  CAS  Google Scholar 

  9. Lagasse E, Connors H, Al-Dhalimy M, Reitsma M, Dohse M, Osborne L, Wang X, Finegold M, Weissman IL, Grompe M (2000) Purified hematopoietic stem cells can differentiate into hepatocytes in vivo. Nat Med 6:1229–1234

    Article  PubMed  CAS  Google Scholar 

  10. Theise ND, Saxena R, Portmann BC, Thung SN, Yee H, Chiriboga L, Kumar A, Crawford JM (1999) The canals of Hering and hepatic stem cells in humans. Hepatology 30:1425–3311

    Article  PubMed  CAS  Google Scholar 

  11. Wang X, Foster M, Al-Dhalimy M, Lagasse E, Finegold M, Grompe M (2003) The origin and liver repopulating capacity of murine oval cells. Proc Natl Acad Sci U S A 100(suppl 1):11881–11888

    Article  PubMed  CAS  Google Scholar 

  12. Nagy P, Evarts RP, McMahon JB, Thorgeirsson SS (1989) Role of TGF-beta in normal differentiation and oncogenesis in rat liver. Mol Carcinog 2:345–354

    Article  PubMed  CAS  Google Scholar 

  13. Couchie D, Holic N, Chobert MN, Corlu A, Laperche Y (2002) In vitro differentiation of WB-F344 rat liver epithelial cells into the biliary lineage. Differentiation (Camb) 69:209–215

    Article  Google Scholar 

  14. Nagai H, Terada K, Watanabe G, Ueno Y, Aiba N, Shibuya T, Kawagoe M, Kameda T, Sato M, Senoo H, Sugiyama T (2002) Differentiation of liver epithelial (stem-like) cells into hepatocytes induced by coculture with hepatic stellate cells. Biochem Biophys Res Commun 293:1420–1425

    Article  PubMed  CAS  Google Scholar 

  15. Miura K, Nagai H, Ueno Y, Goto T, Mikami K, Nakane K, Yoneyama K, Watanabe D, Terada K, Sugiyama T, Imai K, Senoo H, Watanabe S, Watanabe G, Aiba N, Shibuya T, Kawagoe M, Kameda T, Sato M (2003) Epimorphin is involved in differentiation of rat hepatic stem-like cells through cell-cell contact. Differentiation of liver epithelial (stem-like) cells into hepatocytes induced by coculture with hepatic stellate cells. Biochem Biophys Res Commun 311:415–423

    Article  PubMed  CAS  Google Scholar 

  16. Evarts RP, Hu Z, Omori N, Omori M, Marsden ER, Thorgeirsson SS (1996) Precursor-product relationship between oval cells and hepatocytes: comparison between tritiated thymidine and bromodeoxyuridine as tracers. Carcinogenesis (Oxf) 17:2143–2151

    Article  CAS  Google Scholar 

  17. Harlow ELD (1988) Antibodies: a laboratory manual. Cold Spring Harbor Laboratory, New York, pp 139–244

    Google Scholar 

  18. Terada K, Manchikalapudi P, Noiva R, Jauregui HO, Stockert RJ, Schilsky ML (1995) Secretion, surface localization, turnover, and steady state expression of protein disulfide isomerase in rat hepatocytes. J Biol Chem 270:20410–20416

    Article  PubMed  CAS  Google Scholar 

  19. Petersen BE, Goff JP, Greenberger JS, Michalopoulos GK (1998) Hepatic oval cells express the hematopoietic stem cell marker Thy-1 in the rat. Hepatology 27:433–445

    Article  PubMed  CAS  Google Scholar 

  20. Tsao MS, Smith JD, Nelson KG, Grisham JW (1984) A diploid epithelial cell line from normal adult rat liver with phenotypic properties of’ oval’ cells. Exp Cell Res 154:38–52

    Article  PubMed  CAS  Google Scholar 

  21. Blouin R, Blouin MJ, Royal I, Grenier A, Roop DR, Loranger A, Marceau N (1992) Cytokeratin 14 expression in rat liver cells in culture and localization in vivo. Differentiation (Camb) 52:45–54

    Article  CAS  Google Scholar 

  22. Gebhardt R, Schafer-Degenhart I (1988) 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

    Article  PubMed  CAS  Google Scholar 

  23. Dunsford HA, Sell S (1989) Production of monoclonal antibodies to preneoplastic liver cell populations induced by chemical carcinogens in rats and to transplantable Morris hepatomas. Cancer Res 49:4887–4893

    PubMed  CAS  Google Scholar 

  24. Moll R, Franke WW, Schiller DL, Geiger B, Krepler R (1982) The catalog of human cytokeratins: patterns of expression in normal epithelia, tumors and cultured cells. Cell 31:11–24

    Article  PubMed  CAS  Google Scholar 

  25. Moll R, Lowe A, Laufer J, Franke WW (1992) Cytokeratin 20 in human carcinomas. A new histodiagnostic marker detected by monoclonal antibodies. Am J Pathol 140:427–447

    PubMed  CAS  Google Scholar 

  26. Franke WWKJ (1999) Cytokeratins. In: Kreis TVR (ed) Guide book to the cyoskeletal and motor proteins, 2nd edn. Oxford University Press, Oxford

    Google Scholar 

  27. Suzuki A, Zheng Y, Kondo R, Kusakabe M, Takada Y, Fukao K, Nakauchi H, Taniguchi H (2000) Flow-cytometric separation and enrichment of hepatic progenitor cells in the developing mouse liver. Hepatology 32:1230–1239

    Article  PubMed  CAS  Google Scholar 

  28. Hixson DC, Allison JP (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 45:3750–3760

    PubMed  CAS  Google Scholar 

  29. Shiojiri N, Lemire JM, Fausto N (1991) Cell lineages and oval cell progenitors in rat liver development. Cancer Res 51:2611–2620

    PubMed  CAS  Google Scholar 

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Correspondence to Toshihiro Sugiyama.

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Watanabe, G., Nanjo, H., Nagai, H. et al. A novel monoclonal antibody identified hepatic stem-like cells in rats. Med Mol Morphol 44, 103–110 (2011). https://doi.org/10.1007/s00795-010-0516-1

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  • DOI: https://doi.org/10.1007/s00795-010-0516-1

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