Nucleoporation of dendritic cells: efficient gene transfer by electroporation into human monocyte-derived dendritic cells.
Dendritic cells (DCs) are ideal accessory cells in the developing field of gene therapy. Although viral transfection of DCs has become widespread, non-viral transfection of DCs has shown disappointing results. Recently, a new technique for transfecting primary cells has become available -- the Amaxa Nucleofector. Here, we describe the use of this device in the successful non-viral transfection of human monocyte-derived DCs. Using enhanced green fluorescent protein as a reporter gene DCs were transfectable with efficiencies approaching 60%, remaining responsive to lipopolysaccharide-stimulated cytokine production in short-term experiments (though long-term functional assays were hampered by loss of viability). Although these data demonstrate the ease and efficiency with which human monocyte-derived DCs can now be non-virally transfected, they also suggest the limitations of this technology due to the gradual loss of cell viability. The potential use of this system in the development of DC-based cell and gene therapies will be hampered until cell viability can be maintained.
Lenz
Petra
P
Laboratory of Cellular Oncology, National Cancer Institute, National Institutes of Health, Bethesda, MD 20892, USA.
Bacot
Silvia M
SM
Frazier-Jessen
Michelle R
MR
Feldman
Gerald M
GM
D003713:Dendritic Cells / Q000276:immunology / Q000378:metabolism*; D018274:Electroporation; D049452:Green Fluorescent Proteins; D006801:Humans; D016130:Immunophenotyping; D007700:Kinetics; D008164:Luminescent Proteins / Q000378:metabolism; D009000:Monocytes / Q000276:immunology / Q000378:metabolism*; D014162:Transfection
D008164:Luminescent Proteins; D049452:Green Fluorescent Proteins