Nucleoporation of dendritic cells: efficient gene transfer by electroporation into human monocyte‐derived dendritic cells 1

FEBS Letters

Bacot, Silvia M.²  Frazier-Jessen, Michelle R.²  Feldman, Gerald M.²  Lenz, Petra¹ 

[1] Laboratory of Cellular Oncology, National Cancer Institute, NIH, Bethesda, MD 20892, USA;Division of Monoclonal Antibodies, Center for Biologics Evaluation and Research, Food and Drug Administration, HFM-564, Bldg. 29A, Rm 3C24, 29 Lincoln Drive, Bethesda, MD 20892, USA

关键词: Electroporation;    Dendritic cell;    Gene therapy;    Monocyte;    Nucleofection;    Green fluorescent protein;   

DOI  :  10.1016/S0014-5793(03)00169-8

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

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.

【 授权许可】

Unknown