【摘要】 目的:建立从小鼠骨髓中分离、纯化、培养、扩增树突状细胞(DC)前体的方法,研究细胞因子IL-4对小鼠DC体外增殖、分化成熟的影响。方法:将健康小鼠股骨骨髓细胞分离,去除悬浮细胞,将贴壁细胞分为2组,第1组加入重组细胞/巨噬细胞集落刺激因子(GM-CSF)培养;第2组加入GM-CSF和白细胞介素-4(IL-4)联合培养。诱导的DC经相差显微镜观察。结果:分离的DC前体经8~9d的培养,镜下随机取8个视野进行细胞计数,第1组细胞数量是85.38±3.42;第2组细胞数量是119.25±3.96,两组间DC数量有显著差异(t=3.19,P<0.05)。且细胞纯度达90%以上,具有典型的树枝状或裙褶状突起。结论:用GM-CSF和IL-4联合作用更能促进DC的体外扩增及分化。更多还原

【Abstract】 Objective:To establish the methods of isolation,purification and propagation of the dendritic cells(DCs) precursor from the mice bone marrow in vitro and to study the effects of cytokines IL-4 on mice bone marrow DCs.Methods:Bone marrow cells were flushed from the femurs of healthy mice,the floating cells were removed,and the adherent cells were divided into two groups.The Group 1 was continued to be cultured with recombinant granulocyte/macrophage colony stimulating factor(GM-CSF),and the Group 2 was cultured with GM-CSF plus interleukin 4(IL-4).The deduced DCs of two groups were observed by the phase microscope.Results:After DCs were cultured for 8-9 days,8 scopes under the phase microscope.were picked at random and the DC numbers were counted.The DC average number and standard deviation of Group 1 were 85.38 and 3.42,respectively.The DC average number and standard deviation of Group 2 were 119.25 and 3.96,respectively.There was a significant difference between the two groups(t=3.19,P<0.05).The purity of cultured DC was over 90%.The cultured cells in these two groups exhibited special dendritic morphology of the mature DCs.Conclusion:A combination of GM-CSF plus IL-4 can greatly promote the proliferation and differentiation of DCs in vitro.更多还原