两步法从KG-1细胞株体外诱导分化成树突状细胞

Generation of human dendritic cells from KG-1 cells in vitro by two-step process

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 explore the technigue ofgeneration of dendritic cells from human leukemia cell line KG-1 after induction in vitro. Methods： Two-step inductions were carried out： At first,KG-1 cells were treated with Flt3-ligand, TPO and SCF for 5 days. Then the cells were exposed to factors including GH-CSF, IL-4 and TNF-α for 7 days to acquire the properties of mature DC. In day 7,the cells morphology were observed under invert microscopy and the phenotype markets （CD40, CDSO, CD86, HLA-DR）were detected with FACS. Results： After induction with cytokines, KG-1 exhibited morphologic and phenotypic properties of typical dendritic cells. Conclusion： It is possible to generate typical morphological and phenotypic DCs from KG-1 cells by two step process differentiation.