Establishment of dendritomas by fusion of human dendritic cells with human hepatocellular carcinoma cell line HLE cells / 中华肿瘤杂志

ABSTRACT

<p><b>OBJECTIVE</b>To construct dendritomas by fusion of human dendritic cells with HLE cells, a human hepatocellular carcinoma cell line.</p><p><b>METHODS</b>HLE cells were cultured in RPMI 1640 with 15% FCS. Human dendritic cells (DCs) were obtained from peripheral blood monocytes cultured in the presence of GM-CSF and IL-4 for 7 days, matured with TNF-alpha and PGE(2) for 2 days. The DCs and HLE cells were labeled with green fluorescence dye PKH67-GL and red fluorescence dye PKH26-GL, respectively, and fused in 50% polyethylene glycol (PEG) + 10% dimethyl sulfoxide (DMSO) to generate dendritomas for rapid fluorescence-activated cell sorting (FACS).</p><p><b>RESULTS</b>Dendritomas with dual red-green fluorescence were constructed successfully, and FACS analysis showed the effective fusion rate was 16.8%.</p><p><b>CONCLUSION</b>With fluorescence dyes PKH67-GL and PKH26-GL as fusion markers, dendritomas for rapid fluorescence-activated cell sorting are constructed, which may throw new light on immunotherapy of hepatocellular carcinoma.</p>