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目的构建人源树突状细胞(DC)与肝癌细胞系HLE的融合细胞。方法含15%FCS的RPMI1640培养HLE细胞,用GMCSF和IL4培养成人外周血单核细胞7d,并用TNFα和PGE2促成熟2d后获得成熟DC;用荧光染料PKH67GL(绿色荧光)和PKH26GL(红色荧光)分别标记DC和HLE细胞,以50%聚乙二醇和10%二甲基亚砜为融合剂,构建可供流式细胞仪快速筛选的融合细胞。结果成功构建具有红、绿双色荧光的人源DC与HLE的融合细胞,融合率为16.8%。结论利用PKH67GL和PKH26GL为标记,可获得便于快速识别和筛选的DC与肝癌细胞的融合细胞,为使用融合DC疫苗治疗肝癌奠定了基础。
Objective To construct fused cells of human dendritic cells (DCs) and hepatocellular carcinoma cell line HLE. Methods Human peripheral blood mononuclear cells were cultured with RPMI 1640 containing 15% FCS for 7 days, and matured DCs were induced by TNFα and PGE2 for 2 days. Fluorescent dyes PKH67GL (green fluorescence) and PKH26GL (red fluorescence) DC and HLE cells were labeled respectively, and 50% polyethylene glycol and 10% dimethylsulfoxide were used as the fusion reagents to construct fusion cells for rapid screening by flow cytometry. Results The fused cells of human DC with HLE with red and green fluorescence were successfully constructed and the fusion rate was 16.8%. Conclusion Using PKH67GL and PKH26GL as markers, we can obtain fusion cells between DC and hepatoma cells for rapid identification and screening, and lay a foundation for the treatment of hepatocellular carcinoma with DC vaccine.