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目的:探讨采用无血清专用培养基从肺癌患者外周血单核细胞快速培养树突状细胞(DCs)的方法。方法:将10例肺癌患者外周血单核细胞同时采用含人AB型血清培养基和无血清DCs专用培养基(DCMedium)培养,5例于第7天加入促成熟细胞因子组合,第9～10天收获成熟DCs,另5例于第5天加入促成熟细胞因子组合,且TNF-α剂量加大5倍,第7天收获成熟DCs。结果:DCMedium培养的DCs形态变化快,细胞出现突起的时间早;9～10天收获的两种培养基培养的DCs性能无差异;7天收获的DCs中,DCMedium培养的DCs得率和纯度、共刺激分子表达水平、刺激T细胞增殖能力均优于含血清培养基。结论:无血清DCs专用培养基可以在体外快速培养临床级DCs,为肿瘤免疫治疗奠定基础。
Objective: To explore the method of rapidly culturing dendritic cells (DCs) from peripheral blood mononuclear cells of lung cancer patients with serum-free special medium. Methods: Peripheral blood mononuclear cells (PBMCs) from 10 patients with lung cancer were cultured with human AB serum-containing medium and DCMedium without serum. 5 cases were added with mature cytokines on the 7th day, Matured DCs were harvested on day 5, and the other 5 were added to the pro-mature cytokine combination on day 5, with a 5-fold increase in TNF-α dose, and mature DCs were harvested on day 7. Results: DCs cultured in DCMedium showed rapid morphological changes and early protrusion of cells. There was no difference in DCs cultured on the 9th to 10th day. DCs’ yield and purity of DCedium cultured in 7 days, Costimulatory molecules expression levels, stimulate T cell proliferation better than serum-containing medium. Conclusion: Serum-free DCs-specific culture medium can rapidly cultivate clinical DCs in vitro and lay the foundation for tumor immunotherapy.