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目的:探讨两步法从人白血病细胞株(KG-1)体外诱导分化成树突状细胞(DC)的技术。方法:利用两步诱导法,先在培养体系中加入FLT3配体(Flt3-ligand)、血小板生成素(TPO)及干细胞因子(SCF)三种细胞因子,对KG-1细胞进行扩增培养5d;扩增后的细胞在粒细胞-巨噬细胞集落刺激因子(GM-CSF)、白细胞介素4(IL-4)和肿瘤坏死因子(TNF-α)的作用下诱导7d,镜下进行形态学观察并用流式细胞仪分析树突状细胞特征性表面标记CD40、CD80、CD86、HLA-DR的表达情况。结果:诱导7d细胞胞体较大,表面出现许多树突状突起;树突状细胞特征性表面标记CD40、CD80、CD86、HLA-DR表达阳性率显著升高。结论:两步法能促使KG-1细胞分化成具有典型形态和表型特征的DC细胞。
Objective: To investigate the technology of two-step induction of dendritic cells (DC) from human leukemia cell line (KG-1) in vitro. Methods: Three kinds of cytokines, FLT3-ligand, TPO and SCF, were added into the culture system by two-step induction method. KG-1 cells were expanded and cultured for 5 days ; The expanded cells were induced by GM-CSF, IL-4 and TNF-α for 7 days, The expression of CD40, CD80, CD86 and HLA-DR, the characteristic surface markers of dendritic cells, was observed and analyzed by flow cytometry. Results: The somatic cells on the 7th day were large in size with many dendrites on the surface. The positive rates of the characteristic surface markers CD40, CD80, CD86 and HLA-DR on dendritic cells were significantly increased. Conclusion: Two-step method can promote KG-1 cells to differentiate into DC cells with typical morphological and phenotypic characteristics.