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Biological Chemistry

Editor-in-Chief: Brüne, Bernhard

Editorial Board Member: Buchner, Johannes / Ludwig, Stephan / Sies, Helmut / Turk, Boris / Wittinghofer, Alfred

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In vitro differentiation of reprogrammed murine somatic cells into hepatic precursor cells

Tobias Cantz1 / Martina Bleidißel2 / Martin Stehling3 / Hans R. Schöler4

Present address: Junior Research Group Stem Cell Biology, Cluster-of-Excellence ‘REBIRTH’, Hannover Medical School, D-30625 Hannover, Germany
1Department of Cell and Developmental Biology, Max Planck Institute for Molecular Biomedicine, Röntgenstr. 20, D-48149 Münster, Germany

2Department of Cell and Developmental Biology, Max Planck Institute for Molecular Biomedicine, Röntgenstr. 20, D-48149 Münster, Germany

3Department of Cell and Developmental Biology, Max Planck Institute for Molecular Biomedicine, Röntgenstr. 20, D-48149 Münster, Germany

4Department of Cell and Developmental Biology, Max Planck Institute for Molecular Biomedicine, Röntgenstr. 20, D-48149 Münster, Germany

Corresponding author

Citation Information: Biological Chemistry. Volume 389, Issue 7, Pages 889–896, ISSN (Online) 1437-4315, ISSN (Print) 1431-6730, DOI: 10.1515/BC.2008.107, June 2008

Publication History

Received:
2007-11-30
Accepted:
2008-04-23
Published Online:
2008-06-06

Abstract

Recently, a new approach to reprogram somatic cells into pluripotent stem cells was shown by fusion of somatic cells with embryonic stem (ES) cells, which results in a tetraploid karyotype. Normal hepatocytes are often polyploid, so we decided to investigate the differentiation potential of fusion hybrids into hepatic cells. We chose toxic milk mice (a model of Wilson's disease) and performed initial transplantation experiments using this potential cell therapy approach. Mononuclear bone marrow cells from Rosa26 mice were fused with OG2 (Oct4-GFP transgenic) ES cells. Unfused ES cells were eliminated by selection with G418 for OG2-Rosa26 hybrids and fusion-derived colonies could be subcloned. Using an endodermal differentiation protocol, hepatic precursor cells could be generated. After FACS depletion of contaminating Oct4-GFP-positive cells, the hepatic precursor cells were transplanted into immunosuppressed toxic milk mice by intrasplenic injection. However, five out of eight mice showed teratoma formation within 3–6 weeks after transplantation in the spleen and liver. In conclusion, a hepatic precursor cell type was achieved from mononuclear bone marrow cell-ES cell hybrids and preliminary transplantation experiments confirmed engraftment, but also showed teratoma formation, which needs to be excluded by using more stringent purification strategies.

Keywords: embryonic stem cells; endodermal differentiation; fusion; metabolic liver disorders; reprogramming

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