不同浓度粒巨细胞集落刺激因子及白介素-4对树突状细胞体外诱导培养的影响

摘要

Objective To investigate the appropriate concentration range of murine granulocyte-macrophage colony-stimulating factor(GM-CSF)and interleukin-4(IL-4)in incubating mouse bone marrow monocytes-like cells and proper conditions for dendritic cell differentiation.Methods The progenitor of dendritic cells were isolated from bone marrow of Balb/c mice,and incubated in different dose of mouse granuIocyte-macrophage coIony-stimuIating fac-tor and interleukin-4 for continuously seven days,the morphology of cells in each group were observed under micro-scope and snapped with digital camera every other day.The expression of CD1 1 c was detected with flow cytometry. Results Morphological observation revealed that mouse bone marrow monocytes-like cells changed remarkably when cultured with low dose of cytokine.Cells became large and showed dendritic protruding on their surface.Furthermore, most cells were CD1 1 c positive when detected by flow cytometry.The appropriate GM-CSF and IL-4 dose range were 5μg/L~100 μg/L and 2.5 μg/L~50 μg/L respectively,while cytokines of overhigh concentration cause cell death. Conclusion Dendritic cells can be derived from bone marrow monocytes-like cells with appropriate dose of GM-CSF and IL-4 treatment with High purity DCs.%目的：探讨不同浓度小鼠粒/巨噬细胞集落刺激因子（GM-CSF）和白介素-4（IL-4）对小鼠骨髓源性树突状细胞培养的影响，探索树突状细胞诱导培养的适宜条件。方法采用细胞裂解法分离得到Balb/c小鼠骨髓单个核样细胞，分为多组，分别加入不同浓度GM-CSF和IL-4，并设置空白对照组，共培养7 d，每天观察各组细胞形态，隔天拍照。第7天收集细胞，使用流式细胞法检测细胞表面树突状细胞标志分子CD11 c的表达率。结果低浓度GM-CSF及IL-4可培养出高纯度小鼠骨髓源性树突状细胞，高浓度GM-CSF及IL-4可导致细胞死亡，GM-CSF和IL-4的合理细胞诱导浓度范围分别为5～100μg/L和2．5～50μg/L。结论合适浓度的GM-CSF及IL-4可诱导小鼠骨髓单个核样细胞向树突状细胞分化，获得大量树突状细胞。