Ex vivo gene transfer into hepatocytes

Xia Wang, Prashant Mani, Debi P. Sarkar, Namita Roy-Chowdhury, Jayanta Roy-Chowdhury

Research output: Chapter in Book/Report/Conference proceedingChapter

22 Scopus citations

Abstract

Ex vivo gene transfer into hepatocytes could serve several purposes in the context of gene therapy or cell transplantation: (1) isolated hepatocytes can be transduced in culture with therapeutic genes and then transplanted into the recipient; (2) marker genes can be introduced for subsequent identification of transplanted cells and their progeny; (3) gene transfer can be used for conditional immortalization of hepatocytes for expansion in culture; (4) immunomodulatory genes can be transferred into hepatocytes to prevent allograft rejection. Gene transfer into cultured hepatocytes can be achieved using DNA that is not incorporated into recombinant viruses. In such systems, transgene integration into the host cell genome can be enhanced using transposon systems, such as "sleeping beauty." In addition to using the conventional reagents, such as cationic liposomes, DNA transfer into hepatocytes can be achieved by Nucleofection® or special hepatocyte-targeted carriers such as proteoliposomes containing galactose-terminated glycoproteins (e.g. the F protein of the Sendai virus). Alternatively, genes can be transferred using recombinant viruses, such as adenoviral vectors that are episomal or retroviral vectors (including lentiviruses) that permit integration of the transgene into the host genome. Gene transfer using lentiviral vectors has been achieved in both attached and suspended hepatocytes. Transduction efficiency of lentiviral vectors can be enhanced using magnetic nanoparticles (Magnetofection®).

Original languageEnglish (US)
Title of host publicationHepatocyte Transplantation
Subtitle of host publicationMethods and Protocols
PublisherHumana Press
Pages117-139
Number of pages23
ISBN (Print)9781588298836
DOIs
Publication statusPublished - Jan 1 2009

Publication series

NameMethods in Molecular Biology
Volume481
ISSN (Print)1064-3745

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Keywords

  • Ex vivo
  • F-virosome
  • Gene transfer
  • Lentiviral vectors
  • Magnetofection
  • Nucleofection
  • Sleeping beauty

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

Cite this

Wang, X., Mani, P., Sarkar, D. P., Roy-Chowdhury, N., & Roy-Chowdhury, J. (2009). Ex vivo gene transfer into hepatocytes. In Hepatocyte Transplantation: Methods and Protocols (pp. 117-139). (Methods in Molecular Biology; Vol. 481). Humana Press. https://doi.org/10.1007/978-1-59745-201-4_11