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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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
Maggie HW, David AG (2006) Hepatic stem cells: in search of. Stem Cells 24:1833–1840
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
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
Lazaro CA, Rhim JA, Yamada Y, Fausto N (1998) Generation of hepatocytes from oval cell precursors in culture. Cancer Res 58:5514–5522
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
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
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
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
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
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
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
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
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
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
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
Harlow ELD (1988) Antibodies: a laboratory manual. Cold Spring Harbor Laboratory, New York, pp 139–244
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
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
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
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
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
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
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
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
Franke WWKJ (1999) Cytokeratins. In: Kreis TVR (ed) Guide book to the cyoskeletal and motor proteins, 2nd edn. Oxford University Press, Oxford
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
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
Shiojiri N, Lemire JM, Fausto N (1991) Cell lineages and oval cell progenitors in rat liver development. Cancer Res 51:2611–2620
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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