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

E. S. Tzanakakis, D. J. Waxman, L. K. Hansen, Rory P Remmel, Wei-Shou Hu

Research output: Contribution to journalArticle

13 Citations (Scopus)

Abstract

Tissue-like structures of cells organized in vitro have 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 CYP2B1 transgene 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.

Original languageEnglish (US)
Pages (from-to)13-27
Number of pages15
JournalCell Biology and Toxicology
Volume18
Issue number1
DOIs
StatePublished - Apr 13 2002

Fingerprint

Cytochrome P-450 CYP2B1
Gene transfer
Adenoviridae
Rats
Hepatocytes
Genes
Green Fluorescent Proteins
Liver
Apoproteins
Gene encoding
Phenobarbital
Cell culture
Metabolism
Artificial Liver
Dealkylation
Urea
Albumins
Suspensions
Fluorescence
Tissue

Keywords

  • Adenovirus
  • CYP2B
  • GFP
  • Hepatocytes
  • Spheroids

Cite this

Long-term enhancement of cytochrome P450 2B1/2 expression in rat hepatocyte spheroids through adenovirus-mediated gene transfer. / Tzanakakis, E. S.; Waxman, D. J.; Hansen, L. K.; Remmel, Rory P; Hu, Wei-Shou.

In: Cell Biology and Toxicology, Vol. 18, No. 1, 13.04.2002, p. 13-27.

Research output: Contribution to journalArticle

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