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Functional Characterization of Serum-Free Cultured Rat Hepatocytes for Downstream Transplantation Applications

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Abstract:

Although ex vivo culture of hepatocytes is known to impair functionality, it may still be considered as desirable to propagate or manipulate them in culture prior to transplantation into the host liver. The aim of this study was to clarify whether rat hepatocytes cultured over different periods of time proliferate and retain their hepatocyte-specific functions following transplantation into the recipient liver. Rat hepatocytes were cultured under serum-free conditions in the presence of hepatocyte and epidermal growth factors. Cells derived from wild-type donor livers were transplanted into the livers of CD26-deficient rats. Cell proliferation and the expression of hepatocyte-specific markers were determined before and after transplantation. Cell number increased threefold over a culture period of 10 days. The expression of connexin 32 and phosphoenolpyruvate carboxykinase declined over time, indicating the loss of hepatocyte-specific functions. Hepatocytes cultured over 4 or 7 days and then transplanted proliferated in the host parenchyma. The transplanted cells expressed connexin 32, cytokeratin 18, and phosphoenolpyruvate carboxykinase, indicating the differentiated phenotype. The loss of hepatocyte-specific functions during culture may be restored after transplantation, suggesting that the proper physiological environment is required to maintain the differentiated phenotype.

Keywords: Culture; DPPIV; Hepatocyte; Transplantation

Document Type: Research Article

DOI: http://dx.doi.org/10.3727/000000005783982855

Affiliations: 1: Klinik und Poliklinik für Innere Medizin I, Martin-Luther-Universitaet Halle-Wittenberg, D-06097 Halle, Germany, 2: Klinik und Poliklinik für Allgemeinchirurgie, Georg-August-Universitaet Goettingen, D-37075 Goettingen, Germany, 3: Klinik und Poliklinik für Innere Medizin I, Martin-Luther-Universitaet Halle-Wittenberg, D-06097 Halle, Germany 4: Klinik und Poliklinik für Allgemeinchirurgie, Georg-August-Universitaet Goettingen, D-37075 Goettingen, Germany

Publication date: July 1, 2005

More about this publication?
  • Cell Transplantation publishes original, peer-reviewed research and review articles on the subject of cell transplantation and its application to human diseases. To ensure high-quality contributions from all areas of transplantation, separate section editors and editorial boards have been established. Articles deal with a wide range of topics including physiological, medical, preclinical, tissue engineering, and device-oriented aspects of transplantation of nervous system, endocrine, growth factor-secreting, bone marrow, epithelial, endothelial, and genetically engineered cells, among others. Basic clinical studies and immunological research papers are also featured. To provide complete coverage of this revolutionary field, Cell Transplantation will report on relevant technological advances, and ethical and regulatory considerations of cell transplants. Cell Transplantation is now an Open Access journal starting with volume 18 in 2009, and therefore there will be an inexpensive publication charge, which is dependent on the number of pages, in addition to the charge for color figures. This will allow work to be disseminated to a wider audience and also entitle the corresponding author to a free PDF, as well as prepublication of an unedited version of the manuscript.
cog/ct/2005/00000014/00000007/art00009
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