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Identification of a bipotential precursor cell in hepatic cell lines derived from transgenic mice expressing cyto-Met in the liver.

Spagnoli FM, Amicone L, Tripodi M, Weiss MC - J. Cell Biol. (1998)

Bottom Line: Palmate cells show none of these properties.Derivation of epithelial from palmate cells is confirmed by characterization of the progeny of individually fished cells.The clonal isolation of the palmate cell, an immortalized nontransformed bipotential cell that does not yet express the liver-enriched transcription factors and is a precursor of the epithelial-hepatocyte in MMH lines, provides a new tool for the study of mechanisms controlling liver development.

View Article: PubMed Central - PubMed

Affiliation: Unité de Génétique de la Différenciation, URA 1773 du Centre National de la Recherche Scientifique, Institut Pasteur, 75724 Paris Cedex 15, France.

ABSTRACT
Met murine hepatocyte (MMH) lines were established from livers of transgenic mice expressing constitutively active human Met. These lines harbor two cell types: epithelial cells resembling the parental populations and flattened cells with multiple projections and a dispersed growth habit that are designated palmate. Epithelial cells express the liver-enriched transcription factors HNF4 and HNF1alpha, and proteins associated with epithelial cell differentiation. Treatments that modulate their differentiation state, including acidic FGF, induce hepatic functions. Palmate cells show none of these properties. However, they can differentiate along the hepatic cell lineage, giving rise to: (a) epithelial cells that express hepatic transcription factors and are competent to express hepatic functions; (b) bile duct-like structures in three-dimensional Matrigel cultures. Derivation of epithelial from palmate cells is confirmed by characterization of the progeny of individually fished cells. Furthermore, karyotype analysis confirms the direction of the phenotypic transition: palmate cells are diploid and the epithelial cells are hypotetraploid. The clonal isolation of the palmate cell, an immortalized nontransformed bipotential cell that does not yet express the liver-enriched transcription factors and is a precursor of the epithelial-hepatocyte in MMH lines, provides a new tool for the study of mechanisms controlling liver development.

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Characterization of  the epithelial and the palmate cell clone responses to  environmental treatments.  Gelatin, DMSO, and aFGF  treatments lead to epithelial  morphogenesis in the palmate cells. (A and B) Immunofluorescence analysis of  palmate cells treated for 1 wk  with DMSO (A) or aFGF  (B): expression and localization of the epithelial differentiation markers E-cadherin (E-cad.), ZO-1, and  CK, using an anti-pan-CK  antibody. In A, E-cadherin  and the phase-contrast image  are of the same field; in B,  the ZO-1 and the phase-contrast image are of the same  field. (C) Gelatin, DMSO and  aFGF modulate the epithelial  clone towards a differentiated hepatocyte phenotype.  Northern blot analysis of  RNA extracted from the E14  parental line (E14), the epithelial (ep.) and the palmate (pal.) clones as well as control (c) Fao rat hepatoma cells. Cultured cells were examined in the standard growth conditions (−), and 1 wk after growth on gelatin (gel.), or addition of DMSO or aFGF. Each lane was  loaded with 20 μg of total RNA. Probes used are indicated on the left. Bar, 20 μm.
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Figure 6: Characterization of the epithelial and the palmate cell clone responses to environmental treatments. Gelatin, DMSO, and aFGF treatments lead to epithelial morphogenesis in the palmate cells. (A and B) Immunofluorescence analysis of palmate cells treated for 1 wk with DMSO (A) or aFGF (B): expression and localization of the epithelial differentiation markers E-cadherin (E-cad.), ZO-1, and CK, using an anti-pan-CK antibody. In A, E-cadherin and the phase-contrast image are of the same field; in B, the ZO-1 and the phase-contrast image are of the same field. (C) Gelatin, DMSO and aFGF modulate the epithelial clone towards a differentiated hepatocyte phenotype. Northern blot analysis of RNA extracted from the E14 parental line (E14), the epithelial (ep.) and the palmate (pal.) clones as well as control (c) Fao rat hepatoma cells. Cultured cells were examined in the standard growth conditions (−), and 1 wk after growth on gelatin (gel.), or addition of DMSO or aFGF. Each lane was loaded with 20 μg of total RNA. Probes used are indicated on the left. Bar, 20 μm.

Mentions: The growth of the E14 epithelial cells on gelatin-coated (rather than collagen-coated) dishes, or the addition of DMSO or aFGF leads to a more differentiated hepatocyte phenotype. After these treatments, the cells show a more regular shape with more phase dense and granular cytoplasm (Fig. 5). Significantly, they express albumin and transthyretin (TTR, Fig. 6 C). This induced phenotype resembles that presented by the D3 cells in the standard growth conditions.


Identification of a bipotential precursor cell in hepatic cell lines derived from transgenic mice expressing cyto-Met in the liver.

Spagnoli FM, Amicone L, Tripodi M, Weiss MC - J. Cell Biol. (1998)

Characterization of  the epithelial and the palmate cell clone responses to  environmental treatments.  Gelatin, DMSO, and aFGF  treatments lead to epithelial  morphogenesis in the palmate cells. (A and B) Immunofluorescence analysis of  palmate cells treated for 1 wk  with DMSO (A) or aFGF  (B): expression and localization of the epithelial differentiation markers E-cadherin (E-cad.), ZO-1, and  CK, using an anti-pan-CK  antibody. In A, E-cadherin  and the phase-contrast image  are of the same field; in B,  the ZO-1 and the phase-contrast image are of the same  field. (C) Gelatin, DMSO and  aFGF modulate the epithelial  clone towards a differentiated hepatocyte phenotype.  Northern blot analysis of  RNA extracted from the E14  parental line (E14), the epithelial (ep.) and the palmate (pal.) clones as well as control (c) Fao rat hepatoma cells. Cultured cells were examined in the standard growth conditions (−), and 1 wk after growth on gelatin (gel.), or addition of DMSO or aFGF. Each lane was  loaded with 20 μg of total RNA. Probes used are indicated on the left. Bar, 20 μm.
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Related In: Results  -  Collection

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getmorefigures.php?uid=PMC2132947&req=5

Figure 6: Characterization of the epithelial and the palmate cell clone responses to environmental treatments. Gelatin, DMSO, and aFGF treatments lead to epithelial morphogenesis in the palmate cells. (A and B) Immunofluorescence analysis of palmate cells treated for 1 wk with DMSO (A) or aFGF (B): expression and localization of the epithelial differentiation markers E-cadherin (E-cad.), ZO-1, and CK, using an anti-pan-CK antibody. In A, E-cadherin and the phase-contrast image are of the same field; in B, the ZO-1 and the phase-contrast image are of the same field. (C) Gelatin, DMSO and aFGF modulate the epithelial clone towards a differentiated hepatocyte phenotype. Northern blot analysis of RNA extracted from the E14 parental line (E14), the epithelial (ep.) and the palmate (pal.) clones as well as control (c) Fao rat hepatoma cells. Cultured cells were examined in the standard growth conditions (−), and 1 wk after growth on gelatin (gel.), or addition of DMSO or aFGF. Each lane was loaded with 20 μg of total RNA. Probes used are indicated on the left. Bar, 20 μm.
Mentions: The growth of the E14 epithelial cells on gelatin-coated (rather than collagen-coated) dishes, or the addition of DMSO or aFGF leads to a more differentiated hepatocyte phenotype. After these treatments, the cells show a more regular shape with more phase dense and granular cytoplasm (Fig. 5). Significantly, they express albumin and transthyretin (TTR, Fig. 6 C). This induced phenotype resembles that presented by the D3 cells in the standard growth conditions.

Bottom Line: Palmate cells show none of these properties.Derivation of epithelial from palmate cells is confirmed by characterization of the progeny of individually fished cells.The clonal isolation of the palmate cell, an immortalized nontransformed bipotential cell that does not yet express the liver-enriched transcription factors and is a precursor of the epithelial-hepatocyte in MMH lines, provides a new tool for the study of mechanisms controlling liver development.

View Article: PubMed Central - PubMed

Affiliation: Unité de Génétique de la Différenciation, URA 1773 du Centre National de la Recherche Scientifique, Institut Pasteur, 75724 Paris Cedex 15, France.

ABSTRACT
Met murine hepatocyte (MMH) lines were established from livers of transgenic mice expressing constitutively active human Met. These lines harbor two cell types: epithelial cells resembling the parental populations and flattened cells with multiple projections and a dispersed growth habit that are designated palmate. Epithelial cells express the liver-enriched transcription factors HNF4 and HNF1alpha, and proteins associated with epithelial cell differentiation. Treatments that modulate their differentiation state, including acidic FGF, induce hepatic functions. Palmate cells show none of these properties. However, they can differentiate along the hepatic cell lineage, giving rise to: (a) epithelial cells that express hepatic transcription factors and are competent to express hepatic functions; (b) bile duct-like structures in three-dimensional Matrigel cultures. Derivation of epithelial from palmate cells is confirmed by characterization of the progeny of individually fished cells. Furthermore, karyotype analysis confirms the direction of the phenotypic transition: palmate cells are diploid and the epithelial cells are hypotetraploid. The clonal isolation of the palmate cell, an immortalized nontransformed bipotential cell that does not yet express the liver-enriched transcription factors and is a precursor of the epithelial-hepatocyte in MMH lines, provides a new tool for the study of mechanisms controlling liver development.

Show MeSH
Related in: MedlinePlus