Murine Embryonic Stem Cell-Derived Hepatic Progenitor Cells Engraft in Recipient Livers with Limited Capacity of Liver Tissue Formation
Directed endodermal differentiation of murine embryonic stem (ES) cells gives rise to a subset of cells with a hepatic phenotype. Such ES cell-derived hepatic progenitor cells (ES-HPC) can acquire features of hepatocytes in vitro, but fail to form substantial hepatocyte clusters in vivo. In this stu...
| Main Authors: | , , , , , , , , , |
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| Format: | Article |
| Language: | English |
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SAGE Publishing
2008-03-01
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| Series: | Cell Transplantation |
| Online Access: | https://doi.org/10.3727/096368908784153896 |
| _version_ | 1819061531285913600 |
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| author | Amar Deep Sharma Tobias Cantz Arndt Vogel Axel Schambach Dhivya Haridass Markus Iken Martina Bleidißel Michael P. Manns Hans R. Schöler Michael Ott |
| author_facet | Amar Deep Sharma Tobias Cantz Arndt Vogel Axel Schambach Dhivya Haridass Markus Iken Martina Bleidißel Michael P. Manns Hans R. Schöler Michael Ott |
| author_sort | Amar Deep Sharma |
| collection | DOAJ |
| description | Directed endodermal differentiation of murine embryonic stem (ES) cells gives rise to a subset of cells with a hepatic phenotype. Such ES cell-derived hepatic progenitor cells (ES-HPC) can acquire features of hepatocytes in vitro, but fail to form substantial hepatocyte clusters in vivo. In this study, we investigated whether this is due to inefficient engraftment or an immature phenotype of ES-HPC. ES cells engrafted into recipient livers of NOD/SCID mice with a similar efficacy as adult hepatocytes after 28 days. Because transplanted unpurified ES-HPC formed teratomas in the spleen and liver, we applied an albumin promoter/enhancer-driven reporter system to purify ES-HPC by cell sorting. RT-PCR analyses for hepatocyte-specific genes showed that the cells exhibited a hepatic phenotype, lacking the expression of the pluripotency marker Oct4, comparable to cells of day 11.5 embryos. Sorted ES-HPC derived from β-galactosidase transgenic ES cells were injected into fumaryl-acetoacetate-deficient (FAH-/-) SCID mice and analyzed after 8 to 12 weeks. Staining with X-gal solution revealed the presence of engrafted cells throughout the liver. However, immunostaining for the FAH protein indicated hepatocyte formation at a very low frequency, without evidence for large hepatocyte cluster formation. In conclusion, the limited repopulation capacity of ES-HPC is not caused by a failure of primary engraftment, but may be due to an immature hepatic phenotype of the transplanted ES-HPC. |
| first_indexed | 2024-12-21T14:44:22Z |
| format | Article |
| id | doaj.art-a77f46bbb1624820b22c0f3e109a4ed5 |
| institution | Directory Open Access Journal |
| issn | 0963-6897 1555-3892 |
| language | English |
| last_indexed | 2024-12-21T14:44:22Z |
| publishDate | 2008-03-01 |
| publisher | SAGE Publishing |
| record_format | Article |
| series | Cell Transplantation |
| spelling | doaj.art-a77f46bbb1624820b22c0f3e109a4ed52022-12-21T19:00:04ZengSAGE PublishingCell Transplantation0963-68971555-38922008-03-011710.3727/096368908784153896Murine Embryonic Stem Cell-Derived Hepatic Progenitor Cells Engraft in Recipient Livers with Limited Capacity of Liver Tissue FormationAmar Deep Sharma0Tobias Cantz1Arndt Vogel2Axel Schambach3Dhivya Haridass4Markus Iken5Martina Bleidißel6Michael P. Manns7Hans R. Schöler8Michael Ott9Department of Gastroenterology, Hepatology and Endocrinology, Hannover Medical School, Hannover, GermanyDepartment of Cell and Developmental Biology, Max Planck Institute for Molecular Biomedicine, Muenster, GermanyDepartment of Gastroenterology, Hepatology and Endocrinology, Hannover Medical School, Hannover, GermanyDepartment of Experimental Hematology, Hannover Medical School, Hannover, GermanyDepartment of Gastroenterology, Hepatology and Endocrinology, Hannover Medical School, Hannover, GermanyDepartment of Gastroenterology, Hepatology and Endocrinology, Hannover Medical School, Hannover, GermanyDepartment of Cell and Developmental Biology, Max Planck Institute for Molecular Biomedicine, Muenster, GermanyDepartment of Gastroenterology, Hepatology and Endocrinology, Hannover Medical School, Hannover, GermanyDepartment of Cell and Developmental Biology, Max Planck Institute for Molecular Biomedicine, Muenster, GermanyDepartment of Gastroenterology, Hepatology and Endocrinology, Hannover Medical School, Hannover, GermanyDirected endodermal differentiation of murine embryonic stem (ES) cells gives rise to a subset of cells with a hepatic phenotype. Such ES cell-derived hepatic progenitor cells (ES-HPC) can acquire features of hepatocytes in vitro, but fail to form substantial hepatocyte clusters in vivo. In this study, we investigated whether this is due to inefficient engraftment or an immature phenotype of ES-HPC. ES cells engrafted into recipient livers of NOD/SCID mice with a similar efficacy as adult hepatocytes after 28 days. Because transplanted unpurified ES-HPC formed teratomas in the spleen and liver, we applied an albumin promoter/enhancer-driven reporter system to purify ES-HPC by cell sorting. RT-PCR analyses for hepatocyte-specific genes showed that the cells exhibited a hepatic phenotype, lacking the expression of the pluripotency marker Oct4, comparable to cells of day 11.5 embryos. Sorted ES-HPC derived from β-galactosidase transgenic ES cells were injected into fumaryl-acetoacetate-deficient (FAH-/-) SCID mice and analyzed after 8 to 12 weeks. Staining with X-gal solution revealed the presence of engrafted cells throughout the liver. However, immunostaining for the FAH protein indicated hepatocyte formation at a very low frequency, without evidence for large hepatocyte cluster formation. In conclusion, the limited repopulation capacity of ES-HPC is not caused by a failure of primary engraftment, but may be due to an immature hepatic phenotype of the transplanted ES-HPC.https://doi.org/10.3727/096368908784153896 |
| spellingShingle | Amar Deep Sharma Tobias Cantz Arndt Vogel Axel Schambach Dhivya Haridass Markus Iken Martina Bleidißel Michael P. Manns Hans R. Schöler Michael Ott Murine Embryonic Stem Cell-Derived Hepatic Progenitor Cells Engraft in Recipient Livers with Limited Capacity of Liver Tissue Formation Cell Transplantation |
| title | Murine Embryonic Stem Cell-Derived Hepatic Progenitor Cells Engraft in Recipient Livers with Limited Capacity of Liver Tissue Formation |
| title_full | Murine Embryonic Stem Cell-Derived Hepatic Progenitor Cells Engraft in Recipient Livers with Limited Capacity of Liver Tissue Formation |
| title_fullStr | Murine Embryonic Stem Cell-Derived Hepatic Progenitor Cells Engraft in Recipient Livers with Limited Capacity of Liver Tissue Formation |
| title_full_unstemmed | Murine Embryonic Stem Cell-Derived Hepatic Progenitor Cells Engraft in Recipient Livers with Limited Capacity of Liver Tissue Formation |
| title_short | Murine Embryonic Stem Cell-Derived Hepatic Progenitor Cells Engraft in Recipient Livers with Limited Capacity of Liver Tissue Formation |
| title_sort | murine embryonic stem cell derived hepatic progenitor cells engraft in recipient livers with limited capacity of liver tissue formation |
| url | https://doi.org/10.3727/096368908784153896 |
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