Download PDF
Semin Liver Dis 2003; 23(4): 313-324
DOI: 10.1055/s-2004-815558
Copyright © 2003 by Thieme Medical Publishers, Inc., 333 Seventh Avenue, New York, NY 10001, USA. Tel.: +1(212) 584-4662

Isolation and Characterization of Mouse Hepatic Stem Cells in Vitro

Mary C. Weiss1,2 , Hélène Strick-Marchand2
  • 1Professor, Departement de Biologie de Developpement, Institut Pasteur, Paris, France
  • 2Departement de Biologie de Developpement, Institut Pasteur, Paris, France
Further Information

Publication History

Publication Date:
14 January 2004 (online)

    Permissions and Reprints

ABSTRACT

This article covers mouse cell lines showing the properties of hepatoblasts or oval cells and describes hepatocyte differentiation obtained from embryonic stem (ES) cells and mesenchymal adult progenitor cells. It considers the methods used for cultivation and differentiation of the cells, as well as the strengths and weaknesses of the different systems and perspectives for future research. Emphasis is given to hepatoblast cell lines because this is the material for which bipotentiality has been most clearly demonstrated and for which proliferation and differentiation conditions have been worked out.

KEYWORDS

Hepatoblasts - oval cells - hepatocytes - cholangiocytes

REFERENCES

  • 1 Ott M O, Rey-Campos J, Cereghini S, Yaniv M. vHNF1 is expressed in epithelial cells of distinct embryonic origin during development and precedes HNF1 expression.  Mech Dev . 1991;  36 47-58
    CrossrefPubMedGoogle Scholar
  • 2 Nagy P, Bisgaard H C, Thorgeirsson S S. Expression of hepatic transcription factors during liver development and oval cell differentiation.  J Cell Biol . 1994;  126 223-233
    CrossrefPubMedGoogle Scholar
  • 3 Ang S L, Wierda A, Wong D. et al . The formation and maintenance of the definitive endoderm lineage in the mouse: involvement of HNF3/forkhead proteins.  Development . 1993;  119 1301-1315
    PubMedGoogle Scholar
  • 4 Bisgaard H C, Nagy P, Santoni-Rugiu E, Thorgeirsson S S. Proliferation, apoptosis, and induction of hepatic transcription factors are characteristics of the early response of biliary epithelial (oval) cells to chemical carcinogens.  Hepatology . 1996;  23 62-70
    PubMedGoogle Scholar
  • 5 Rausa F, Samadani U, Ye H. et al . The cut-homeodomain transcriptional activator HNF-6 is coexpressed with its target gene HNF-3 beta in the developing murine liver and pancreas.  Dev Biol . 1997;  192 228-246
    CrossrefPubMedGoogle Scholar
  • 6 Rausa F M, Galarneau L, Belanger L, Costa R H. The nuclear receptor fetoprotein transcription factor is coexpressed with its target gene HNF-3beta in the developing murine liver, intestine and pancreas.  Mech Dev . 1999;  89 185-188
    CrossrefPubMedGoogle Scholar
  • 7 Duncan S A, Manova K, Chen W S. et al . Expression of transcription factor HNF-4 in the extraembryonic endoderm, gut, and nephrogenic tissue of the developing mouse embryo: HNF-4 is a marker for primary endoderm in the implanting blastocyst.  Proc Natl Acad Sci USA . 1994;  91 7598-7602
    PubMedGoogle Scholar
  • 8 Kuo C F, Xanthopoulos K G, Darnell J E. Fetal and adult localization of C/EBP: evidence for combinatorial action of transcription factors in cell-specific gene expression.  Development . 1990;  109 473-481
    PubMedGoogle Scholar
  • 9 Barbacci E, Reber M, Ott M O. et al . Variant hepatocyte nuclear factor 1 is required for visceral endoderm specification.  Development . 1999;  126 4795-4805
    PubMedGoogle Scholar
  • 10 Coffinier C, Thépot D, Babinet C, Yaniv M, Barra J. Essential role for the homeoprotein vHNF1/HNF1B in visceral endoderm differentiation.  Development . 1999;  126 4785-4794
    PubMedGoogle Scholar
  • 11 Coffinier C, Gresh L, Fiette L. et al . Bile system morphogenesis defects and liver dysfunction upon targeted deletion of HNF1β.  Development . 2002;  129 1829-1838
    PubMedGoogle Scholar
  • 12 Landry C, Clotman F, Hioki T. et al . HNF-6 is expressed in endoderm derivatives and nervous system of the mouse embryo and participates to the cross-regulatory network of liver-enriched transcription factors.  Dev Biol . 1997;  192 247-257
    CrossrefPubMedGoogle Scholar
  • 13 Clotman F, Lannoy V J, Reber M. et al . The onecut transcription factor HNF6 is required for normal development of the biliary tract.  Development . 2002;  129 1819-1828
    PubMedGoogle Scholar
  • 14 Germain L, Blouin M J, Marceau N. Biliary epithelial and hepatocytic cell lineage relationships in embryonic rat liver as determined by the differential expression of cytokeratins, a-fetoprotein, albumin, and cell surface-exposed components.  Cancer Res . 1988;  48 4909-4918
    PubMedGoogle Scholar
  • 15 Shiojiri N, Lemire J M, Fausto N. Cell lineages and oval cell progenitors in rat liver development.  Cancer Res . 1991;  51 2611-2620
    PubMedGoogle Scholar
  • 16 Petersen B E, Goff J P, Greenberger J S, Michalopoulos G K. Hepatic oval cells express the hematopoietic stem cell marker Thy-1 in the rat.  Hepatology . 1998;  27 433-445
    CrossrefPubMedGoogle Scholar
  • 17 Omori N, Omori M, Evarts R P. et al . Partial cloning of rat CD34 cDNA and expression during stem cell-dependent liver regeneration in the adult rat.  Hepatology . 1997;  26 720-727
    PubMedGoogle Scholar
  • 18 Fujio K, Hu Z, Evarts R P. et al . Coexpression of stem cell factor and c-kit in embryonic and adult liver.  Exp Cell Res . 1996;  224 243-250
    CrossrefPubMedGoogle Scholar
  • 19 Zhang M, Thorgeirsson S S. Modulation of connexins during differentiation of oval cells into hepatocytes.  Exp Cell Res . 1994;  213 37-42
    CrossrefPubMedGoogle Scholar
  • 20 Volpes R, van den Oord J J, Desmet V J. Integrins as differential cell lineage markers of primary liver tumors.  Am J Pathol . 1993;  142 1483-1492
    PubMedGoogle Scholar
  • 21 Couvelard A, Bringuier A F, Dauge M C. et al . Expression of integrins during liver organogenesis in humans.  Hepatology . 1998;  27 839-847
    CrossrefPubMedGoogle Scholar
  • 22 Rogler L E. Selective bipotential differentiation of mouse embryonic hepatoblasts in vitro.  Am J Pathol . 1997;  150 591-602
    PubMedGoogle Scholar
  • 23 Glickman R M, Sabesin S M. Lipoprotein metabolism. In: Arias IM, Boyer JL, Jakoby WB, et al, eds. The Liver Biology and Pathobiology New York: Raven Press 1994: 391-415
    Google Scholar
  • 24 Holic N, Suzuki T, Corlu A. et al . Differential expression of the rat gamma-glutamyl transpeptidase gene promoters along with differentiation of hepatoblasts into biliary or hepatocytic lineage.  Am J Pathol . 2000;  157 537-548
    PubMedGoogle Scholar
  • 25 Tietjen T G, Mjaatvedt C H, Yang V W. Expression of class I alcohol dehydrogenase gene in developing rat fetuses.  J Histochem Cytochem . 1994;  42 745-753
    PubMedGoogle Scholar
  • 26 Vonesch J L, Nakshatri H, Philippe M, Chambon P, Dolle P. Stage and tissue-specific expression of the alcohol dehydrogenase 1 (Adh-1) gene during mouse development.  Dev Dyn . 1994;  199 199-213
    PubMedGoogle Scholar
  • 27 Guillouzo A, Weber A, Le Provost E, Rissel M, Schapira F. Cell types involved in the expression of foetal aldolases during rat azo-dye hepatocarcinogenesis.  J Cell Sci . 1981;  49 249-260
    PubMedGoogle Scholar
  • 28 Numazaki M, Tsutsumi K, Tsutsumi R, Ishikawa K. Expression of aldolase isozyme mRNAs in fetal rat liver.  Eur J Biochem . 1984;  142 165-170
    CrossrefPubMedGoogle Scholar
  • 29 Petersen B E, Bowen W C, Patrene K D. et al . Bone marrow as a potential source of hepatic oval cells.  Science . 1999;  284 1168-1170
    CrossrefPubMedGoogle Scholar
  • 30 Theise N D, Nimmakayalu M, Gardner R. et al . Liver from bone marrow in humans.  Hepatology . 2000;  32 11-16
    CrossrefPubMedGoogle Scholar
  • 31 Lagasse E, Connors H, Al-Dhalimy M. et al . Purified hematopoietic stem cells can differentiate into hepatocytes in vivo.  Nat Med . 2000;  6 1229-1234
    CrossrefPubMedGoogle Scholar
  • 32 Wang X, Al-Dhalimy M, Lagasse E, Finegold M, Grompe M. Liver repopulation and correction of metabolic liver disease by transplanted adult mouse pancreatic cells.  Am J Pathol . 2001;  158 571-579
    CrossrefPubMedGoogle Scholar
  • 33 Schwartz R E, Reyes M, Koodie L. et al . Multipotent adult progenitor cells from bone marrow differentiate into functional hepatocyte-like cells.  J Clin Invest . 2002;  109 1291-1302
    CrossrefPubMedGoogle Scholar
  • 34 Jiang Y, Jahagirdar B N, Reinhardt R L. et al . Pluripotency of mesenchymal stem cells derived from adult marrow.  Nature . 2002;  418 41-49
    CrossrefPubMedGoogle Scholar
  • 34a Wang X, Willenbring H, Akkari Y. et al . Cell fusion is the principal source of bone-marrow-derived hepatocytes.  Nature . 2003;  422 897-901
    CrossrefPubMedGoogle Scholar
  • 34b Vassilopoulos G, Wang P R, Russell D W. Transplanted bone marrow regenerates liver by cell fusion.  Nature . 2003;  422 901-904
    CrossrefPubMedGoogle Scholar
  • 35 Zheng Y-W, Suzuki A, Taniguchi H. Diversity of hepatic stem cells in the fetal and adult liver.  Semin Liver Dis . 2003;  23 xxx
    PubMedGoogle Scholar
  • 36 Nitou M, Sugiyama Y, Ishikawa K, Shiojiri N. Purification of fetal mouse hepatoblasts by magnetic beads coated with monoclonal anti-E-cadherin antibodies and their in vitro culture.  Exp Cell Res . 2002;  279 330-343
    CrossrefPubMedGoogle Scholar
  • 37 Suzuki A, Zheng Y W, Kondo R. et al . Flow-cytometric separation and enrichment of hepatic progenitor cells in the developing mouse liver.  Hepatology . 2000;  32 1230-1239
    CrossrefPubMedGoogle Scholar
  • 38 Suzuki A, Zheng Y W, Kaneko S. et al . Clonal identification and characterization of self-renewing pluripotent stem cells in the developing liver.  J Cell Biol . 2002;  156 173-184
    CrossrefPubMedGoogle Scholar
  • 39 Amicone L, Spagnoli F M, Späth G. et al . Transgenic expression in the liver of truncated Met blocks apoptosis and permits immortalization of hepatocytes.  EMBO J . 1997;  16 495-503
    CrossrefPubMedGoogle Scholar
  • 40 Spagnoli F M, Amicone L, Tripodi M, Weiss M C. Identification of a bipotential precursor cell in hepatic cell lines derived from transgenic mice expressing cyto-Met in the liver.  J Cell Biol . 1998;  143 1101-1112
    CrossrefPubMedGoogle Scholar
  • 41 Spagnoli F M, Cicchini C, Tripodi M, Weiss M C. Inhibition of MMH (Met murine hepatocyte) cell differentiation by TGF(beta) is abrogated by pre-treatment with the heritable differentiation effector FGF1.  J Cell Sci . 2000;  113 3639-3647
    PubMedGoogle Scholar
  • 42 Aiuti A, Cicchini C, Bernardini S. et al . Hematopoietic support and cytokine expression of murine-stable hepatocyte cell lines (MMH).  Hepatology . 1998;  28 1645-1654
    CrossrefPubMedGoogle Scholar
  • 43 Chagraoui J, Lepage-Noll A, Anjo A, Uzan G, Charbord P. Fetal liver stroma consists of cells in epithelial-to-mesenchymal transition.  Blood . 2003;  101 2973-2982
    CrossrefPubMedGoogle Scholar
  • 44 Strick-Marchand H, Weiss M C. Inducible differentiation and morphogenesis of bipotential liver cell lines from wild-type mouse embryos.  Hepatology . 2002;  36 794-805
    PubMedGoogle Scholar
  • 45 Mennone A, Alvaro D, Cho W, Boyer J L. Isolation of small polarized bile duct units.  Proc Natl Acad Sci USA . 1995;  92 6527-6531
    PubMedGoogle Scholar
  • 46 Cho W K, Mennone A, Boyer J L. Isolation of functional polarized bile duct units from mouse liver.  Am J Physiol Gastrointest Liver Physiol . 2001;  280 G241-G246
    PubMedGoogle Scholar
  • 47 Strick-Marchand H, Weiss M C. Embryonic liver cells and permanent lines as models for hepatocyte and bile duct cell differentiation.  Mech Dev . 2003;  120 89-99
    CrossrefPubMedGoogle Scholar
  • 48 Coleman W B, Grisham J W. Epithelial stem-like cells of the rodent liver. In: Strain AJ, Diehl AM, eds. Liver Growth and Repair London: Chapman & Hall 1998: 50-99
    Google Scholar
  • 49 Fausto N, Campbell J S. The role of hepatocytes and oval cells in liver regeneration and repopulation.  Mechanisms of Development . 2003;  120 117-130
    CrossrefPubMedGoogle Scholar
  • 50 Richards W G, Yoder B K, Isfort R J. et al . Isolation and characterization of liver epithelial cell lines from wild-type and mutant TgN737Rpw mice.  Am J Pathol . 1997;  150 1189-1197
    PubMedGoogle Scholar
  • 51 Richards W G, Yoder B K, Isfort R J. et al . Oval cell proliferation associated with the murine insertional mutation TgN737Rpw.  Am J Pathol . 1996;  149 1919-1930
    PubMedGoogle Scholar
  • 52 Dumble M L, Croager E J, Yeoh G CT, Quail E A. Generation and characterization of p53 null transformed hepatic progenitor cells: oval cells give rise to hepatocellular carcinoma.  Carcinogenesis . 2002;  23 435-445
    CrossrefPubMedGoogle Scholar
  • 53 Petersen B E, Grossbard B, Hatch H, Pi L, Deng J, Scott E W. Mouse A6-positive hepatic oval cells also express several hematopoietic stem cell markers.  Hepatology . 2003;  37 632-640
    CrossrefPubMedGoogle Scholar
  • 54 Hamazaki T, Iiboshi Y, Oka M. et al . Hepatic maturation in differentiating embryonic stem cells in vitro.  FEBS Lett . 2001;  497 15-19
    CrossrefPubMedGoogle Scholar
  • 55 Miyashita H, Suzuki A, Fukao K, Nakauchi H, Taniguchi H. Evidence for hepatocyte differentiation from embryonic stem cells in vitro.  Cell Transplant . 2002;  11 429-434
    PubMedGoogle Scholar
  • 56 Chinzei R, Tanaka Y, Shimizu-Saito K. et al . Embryoid-body cells derived from a mouse embryonic stem cell lines show differentiation into functional hepatocytes.  Hepatology . 2002;  36 22-29
    CrossrefPubMedGoogle Scholar
  • 57 Ishizaka S, Shiori A, Kanda S. et al . Development of hepatocytes from embryonic stem cells after transfection with the HNF3beta gene.  FASEB J . 2002;  16 1444-1454
    PubMedGoogle Scholar
  • 58 Reyes M, Lund T, Lenvik T. et al . Purification and ex vivo expansion of post-natal human marrow mesodermal progenitor cells.  Blood . 2001;  98 2615-2625
    CrossrefPubMedGoogle Scholar
  • 59 Ivanova N B, Dimos J T, Schaniel C. et al . A stem cell molecular signature.  Science . 2002;  298 601-604
    CrossrefPubMedGoogle Scholar
  • 60 Ramalho-Santos M, Yoon S, Matsuzaki Y, Mulligan R C, Melton D A. “Stemness”: transcriptional profiling of embryonic and adult stem cells.  Science . 2002;  298 597-600
    CrossrefPubMedGoogle Scholar
  • 61 Plescia C, Rogler C, Rogler L. Genomic expression analysis implicates Wnt signaling pathway and extracellular matrix alterations in hepatic specification and differentiation of murine hepatic stem cells.  Differentiation . 2001;  68 254-269
    CrossrefPubMedGoogle Scholar
  • 62 Wu J C, Merlino G, Cveklova K, Mosinger Jr B, Fausto N. Autonomous growth in serum-free medium and production of hepatocellular carcinomas by differentiated hepatocyte lines that overexpress transforming growth factor alpha 1.  Cancer Res . 1994;  54 5964-5973
    PubMedGoogle Scholar
  • 63 Wu J C, Merlino G, Fausto N. Establishment and characterization of differentiated, nontransformed hepatocyte cell lines derived from mice transgenic for transforming growth factor α.  Proc Natl Acad Sci USA . 1994;  91 674-678
    PubMedGoogle Scholar
  • 64 Matise M P, Auerbach W, Joyner A L. Production of targeted embryonic stem cell clones. In: Joyner AL, ed. Gene Targeting: a Practical Approach Oxford: Oxford University Press 1999: 101-131
    Google Scholar
  • 65 Tang D G, Tokumoto Y M, Apperly J A, Lloyd A C, Raff M C. Lack of replicative senescence in cultured rat oligodendrocyte precursor cells.  Science . 2001;  291 868-871
    CrossrefPubMedGoogle Scholar
  • 66 Mathon N F, Malcolm D S, Harrisingh M C, Cheng L, Lloyd A C. Lack of replicative senescence in normal rodent glia.  Science . 2001;  291 868-875
    CrossrefPubMedGoogle Scholar
  • 67 Ramirez R D, Morales C P, Herbert B S. et al . Putative telomere-independent mechanisms of replicative aging reflect inadequate growth conditions.  Genes Dev . 2001;  15 398-403
    CrossrefPubMedGoogle Scholar
      >