Long-term enhancement of cytochrome P450 2B1/2 expression in rat hepatocyte spheroids through adenovirus-mediated gene transfer |
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Authors: | E.S. Tzanakakis D.J. Waxman L.K. Hansen R.P. Remmel W.-S. Hu |
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Affiliation: | (1) Department of Chemical Engineering and Materials Science, University of Minnesota, Minneapolis, Minnesota, USA;(2) Department of Biology, Division of Cell and Molecular Biology, Boston University, Boston, Massachusetts, USA;(3) Department of Laboratory Medicine and Pathology, University of Minnesota, Minneapolis, Minnesota, USA;(4) Department of Medicinal Chemistry, University of Minnesota, Minneapolis, Minnesota, USA |
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Abstract: | Tissue-like structures of cells organized in vitrohave a great potential for a number of clinical and biomedical applications. Cell functions may be modulated with gene delivery, improving the characteristics of these structures. Hepatocytes that self-assemble into spheroids can be transduced through adenovirus-mediated gene transfer. An adenoviral vector (AdGFP) was employed to deliver a gene encoding for green fluorescent protein (GFP) in rat hepatocyte spheroids. GFP fluorescence was detected for at least one month. Furthermore, the rat cytochrome P450 2B1 gene (CYP2B1) was transferred through infection with a recombinant adenovirus (AdCYP2B1) in hepatocyte spheroids cultured in suspension. The CYP2B1/2 mRNA and apoprotein levels were continuously higher for over 23 days compared to phenobarbital-induced and control cultures. P450-catalyzed pentoxyresorufin-O-dealkylation activity was also high in the AdCYP2B1-infected spheroids. In these spheroid cultures, albumin and urea levels were similar to those in uninfected spheroid cultures, indicating that expression of the CYP2B1transgene did not impair these liver-specific functions. Hepatocyte spheroids transduced by recombinant adenoviral vectors can be efficiently used for drug metabolism studies, in implantation, and in bioartificial liver devices. This revised version was published online in August 2006 with corrections to the Cover Date. |
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Keywords: | adenovirus CYP2B GFP hepatocytes spheroids |
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