Primary human hepatocytes on biodegradable poly(l-lactic acid) matrices: A promising model for improving transplantation efficiency with tissue engineering

Authors

  • Eva Török,

    1. Departments of Hepatobiliary and Transplant Surgery, University Medical Center Hamburg-Eppendorf, Hamburg, Germany
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    • *These authors contributed equally to this work.

  • Marc Lutgehetmann,

    1. Departments of Internal Medicine I, University Medical Center Hamburg-Eppendorf, Hamburg, Germany
    2. Departments of Medical Microbiology,Virology, and Hygiene, University Medical Center Hamburg-Eppendorf, Hamburg, Germany
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    • *These authors contributed equally to this work.

  • Jeanette Bierwolf,

    1. Departments of Hepatobiliary and Transplant Surgery, University Medical Center Hamburg-Eppendorf, Hamburg, Germany
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  • Stefan Melbeck,

    1. Departments of Hepatobiliary and Transplant Surgery, University Medical Center Hamburg-Eppendorf, Hamburg, Germany
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  • Jochen Düllmann,

    1. Departments of Anatomy II: Experimental Morphology, University Medical Center Hamburg-Eppendorf, Hamburg, Germany
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  • Bjoern Nashan,

    1. Departments of Hepatobiliary and Transplant Surgery, University Medical Center Hamburg-Eppendorf, Hamburg, Germany
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  • Peter X. Ma,

    1. Department of Biologic and Materials Sciences, University of Michigan, Ann Arbor, MI
    2. Macromolecular Science and Engineering Center, University of Michigan, Ann Arbor, MI
    3. Department of Biomedical Engineering, University of Michigan, Ann Arbor, MI
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  • Joerg M. Pollok

    Corresponding author
    1. Departments of Hepatobiliary and Transplant Surgery, University Medical Center Hamburg-Eppendorf, Hamburg, Germany
    • Department of Hepatobiliary and Transplant Surgery, University Medical Center Hamburg-Eppendorf, Martinistrasse 52, 20246 Hamburg, Germany
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    • Telephone: +49-40-7410-58572; FAX: +49-40-7410-45311


Abstract

Liver transplantation is an established treatment for acute and chronic liver disease. However, because of the shortage of donor organs, it does not fulfill the needs of all patients. Hepatocyte transplantation is promising as an alternative method for the treatment of end-stage liver disease and as bridging therapy until liver transplantation. Our group has been working on the optimization of matrix-based hepatocyte transplantation. In order to increase cell survival after transplantation, freshly isolated human hepatocytes were seeded onto biodegradable poly(l-lactic acid) (PLLA) polymer scaffolds and were cultured in a flow bioreactor. PLLA discs were seeded with human hepatocytes and exposed to a recirculated medium flow for 6 days. Human hepatocytes formed spheroidal aggregates with a liver-like morphology and active metabolic function. Phase contrast microscopy showed increasing numbers of spheroids of increasing diameter during the culture period. Hematoxylin and eosin histology showed viable and intact hepatocytes inside the spheroids. Immunohistochemistry confirmed sustained hepatocyte function and a preserved hepatocyte-specific cytoskeleton. Albumin, alpha-1-antitrypsin, and urea assays showed continued production during the culture period. Northern blot analysis demonstrated increasing albumin signals. Scanning electron micrographs showed hepatocyte spheroids with relatively smooth undulating surfaces and numerous microvilli. Transmission electron micrographs revealed intact hepatocytes and junctional complexes with coated pits and vesicles inside the spheroids. Therefore, we conclude that primary human hepatocytes, precultured in a flow bioreactor on a PLLA scaffold, reorganize to form morphologically intact liver neotissue, and this might offer an optimized method for hepatocyte transplantation because of the expected reduction of the initial cell loss, the high regenerative potential in vivo, and the preformed functional integrity. Liver Transpl 17:104–114, 2011. © 2011 AASLD.

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