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  

Generation of dendritic cells with appropriate concentration of granuIocyte-macrophage coIony-stimuIating factor and interleukin-4

作　　者：王晓婧 梅晓冬 

机构地区：[1]安徽医科大学附属省立医院,安徽省立医院呼吸内科,合肥230001

出　　处：《中国临床保健杂志》2014年第2期157-159,I0001,共4页Chinese Journal of Clinical Healthcare

基　　金：2013年度安徽省自然科学基金项目(1308085MH115)

摘　　要：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CD11c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。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＆amp;nbsp;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.

关 键 词：树突细胞 细胞培养技术 巨噬细胞集落刺激因子 白细胞介素4 INTERLEUKIN-4 

分 类 号：R392[医药卫生—免疫学]