目的:建立自体热休克凋亡肺癌细胞抗原制备、树突状细胞(DC)体外诱导、以及抗原负载方法,为制备DC肿瘤疫苗提供技术基础。方法:采用酶消化法从手术切除的肺癌新鲜组织获得单细胞悬液,热休克后用白桦脂酸诱导其凋亡制备成细胞抗原;采用血细胞分离机采集外周血单个核细胞(PBMC),贴壁法获得单核细胞,经GM-CSF与IL-4体外诱导成未成熟树突状细胞(imDC);负载细胞抗原后制备成DC肿瘤疫苗,并对DC疫苗的形态、数量及表型特征进行分析。结果:(1)肿瘤细胞抗原得率:(13.68±1.30)×106/g组织;平均凋亡率:(93.79±2.31)%;(2)imDC平均得率为(9.37±0.83)×106/(118.77±12.56)×106个PBMC,活率>95%;imDC表型分析:CD11c+CD14-、CD11c+HLA-DR+、CD11c+CD80+、CD11c+CD83+、CD11c+CD86+表达率分别为(87.45±3.42)%、(87.16±1.08)%、(2.80±1.52)%、(5.64±1.56)%、(5.11±1.09)%;(3)DC疫苗平均得率为(5.75±0.69)×106/(9.37±0.83)×106个imDC,活率>95%,DC疫苗表型:CD11c+CD14-、CD11c+HLA-DR+、CD11c+CD80+、CD11c+CD83+、CD11c+CD86+表达率分别为(92.73±1.21)%、(89.35±2.31)%、(86.80±1.23)%、(69.53±7.21)%、(94.92±1.48)%。结论:该方法稳定、安全、可靠,可制备出具有成熟DC表型的肿瘤疫苗。 OBJECTIVE: To establish the preparation of anti-apoptotic lung cancer cells in vitro, the induction of dendritic cells (DCs) in vitro, and the method of antigen loading, providing the technical basis for the preparation of DC tumor vaccines. Methods: The single cell suspension was obtained from freshly resected lung cancer tissues by enzymatic digestion method. After heat shock, the cells were induced to apoptosis by lipids, then the peripheral blood mononuclear cells (PBMCs) were collected by using a blood cell separator. Monocytes were obtained by wall-method, immature dendritic cells (imDC) were induced by GM-CSF and IL-4 in vitro. DC vaccines were prepared after loading cell antigens. The morphology, quantity and phenotypic characteristics Analyze. The average rate of apoptosis was (93.79 ± 2.31)%; (2) The average yield of imDC was (9.37 ± 0.83) × 106 / g, (118.77 ± 12.56) × 106 PBMCs, and the viability was> 95%. The imDC phenotype analysis showed that the expression rates of CD11c + CD14-, CD11c + HLA-DR +, CD11c + CD80 +, CD11c + CD83 + and CD11c + CD86 + were (87.45 ± 3.42%, 87.16 ± 1.08%, 2.80 ± 1.52%, 5.64 ± 1.56% and 5.11 ± 1.09% respectively. (3) The average yield of DC vaccine was (5.75 ± 0.69) × 106 / ( 9.37 ± 0.83) × 106 imDC, and the viability> 95%. The DC vaccine phenotypes were (92.73 ± 1.21), CD11c + CD14-, CD11c + HLA-DR +, CD11c + CD80 +, CD11c + CD83 + and CD11c + ), (89.35 ± 2.31)%, (86.80 ± 1.23)%, (69.53 ± 7.21)% and (94.92 ± 1.48)%, respectively. Conclusion: The method is stable, safe and reliable, and can produce tumor vaccine with mature DC phenotype.