title: Ex vivo gene transfer into hepatocytes.
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).
authors.affiliation: Department of Medicine and Molecular Genetics, and the Marion Bessin Liver Research Center, Albert Einstein College of Medicine, New York, USA.
authors.forename: Xia
authors.initials: X
authors.lastname: Wang
authors.forename: Prashant
authors.initials: P
authors.lastname: Mani
authors.forename: Debi P
authors.initials: DP
authors.lastname: Sarkar
authors.forename: Namita
authors.initials: N
authors.lastname: Roy-Chowdhury
authors.forename: Jayanta
authors.initials: J
authors.lastname: Roy-Chowdhury
mesh_terms: D000818:Animals; D002461:Cell Line, Transformed / Q000166:cytology; D018014:Gene Transfer Techniques; D015316:Genetic Therapy / Q000379:methods; D022781:Hepatocytes / Q000166:cytology / Q000378:metabolism* / Q000637:transplantation; D006801:Humans; D016031:Liver Transplantation / Q000379:methods; D008280:Magnetics / Q000379:methods; D008954:Models, Biological; D053758:Nanoparticles / Q000627:therapeutic use; D013194:Staining and Labeling / Q000379:methods; D019172:Transplantation Conditioning / Q000379:methods