摘要
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 dendritomas by fusion of human dendritic cells with HLE cells, a human hepatocellular carcinoma cell line. Methods 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-α and PGE2 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 fluoreseence-activated cell sorting （FACS）. Results Dendritomas with dual red-green fluorescence were constructed successfully, and FACS analysis showed the effective fusion rate was 16.8%. Condttsion 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.
出处
《中华肿瘤杂志》
CAS
CSCD
北大核心
2005年第8期465-467,共3页
Chinese Journal of Oncology
基金
卫生部部属(管)医疗机构临床学科重点资助项目(2001)
