Long-term enhancement of cytochrome P450 2B1/2 expression in rat hepatocyte spheroids through adenovirus-mediated gene transfer View Full Text


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Article Info

DATE

2002-02

AUTHORS

E.S. Tzanakakis, D.J. Waxman, L.K. Hansen, R.P. Remmel, W.-S. Hu

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. More... »

PAGES

13-27

References to SciGraph publications

  • 1991-04-22. Manipulation of Adenovirus Vectors in GENE TRANSFER AND EXPRESSION PROTOCOLS
  • 1992-08. Adenovirus–mediated in vivo gene transfer and expression in normal rat liver in NATURE GENETICS
  • 1995-05. Extended liver-specific functions of porcine hepatocyte spheroids entrapped in collagen gel in IN VITRO CELLULAR & DEVELOPMENTAL BIOLOGY - ANIMAL
  • 1996-04. Mechanistics of formation and ultrastructural evaluation of hepatocyte spheroids in IN VITRO CELLULAR & DEVELOPMENTAL BIOLOGY - ANIMAL
  • 1993-12. Adenovirus mediated expression of therapeutic plasma levels of human factor IX in mice in NATURE GENETICS
  • 1992-05. Gene transfer in primary cultures of human hepatocytes in IN VITRO CELLULAR & DEVELOPMENTAL BIOLOGY
  • Identifiers

    URI

    http://scigraph.springernature.com/pub.10.1023/a:1014415128392

    DOI

    http://dx.doi.org/10.1023/a:1014415128392

    DIMENSIONS

    https://app.dimensions.ai/details/publication/pub.1009976291

    PUBMED

    https://www.ncbi.nlm.nih.gov/pubmed/11991083


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