Ex vivo gene transfer into hepatocytes.

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

Research output: Contribution to journalArticle

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)
Pages (from-to)117-140
Number of pages24
JournalMethods in molecular biology (Clifton, N.J.)
Volume481
StatePublished - 2009

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Hepatocytes
Genes
Transgenes
Viral Fusion Proteins
Genome
Viruses
Systems Integration
Beauty
Lentivirus
DNA
Cell Transplantation
Galactose
Liposomes
Genetic Therapy
Nanoparticles
Allografts
Glycoproteins

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

Cite this

Ex vivo gene transfer into hepatocytes. / Wang, Xia; Mani, Prashant; Sarkar, Debi P.; Roy-Chowdhury, Namita; Roy-Chowdhury, Jayanta.

In: Methods in molecular biology (Clifton, N.J.), Vol. 481, 2009, p. 117-140.

Research output: Contribution to journalArticle

Wang, X, Mani, P, Sarkar, DP, Roy-Chowdhury, N & Roy-Chowdhury, J 2009, 'Ex vivo gene transfer into hepatocytes.', Methods in molecular biology (Clifton, N.J.), vol. 481, pp. 117-140.
Wang, Xia ; Mani, Prashant ; Sarkar, Debi P. ; Roy-Chowdhury, Namita ; Roy-Chowdhury, Jayanta. / Ex vivo gene transfer into hepatocytes. In: Methods in molecular biology (Clifton, N.J.). 2009 ; Vol. 481. pp. 117-140.
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