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[目的]通过比较常用的M-CSF与L929细胞培养上清两种诱导方式获得骨髓巨噬细胞(BMDM)的形态及基本生物学功能,为选择制备BMDM的方法提供实验依据。[方法]用两种方式诱导小鼠骨髓获取BMDM,然后分别利用Zeiss荧光显微镜和流式细胞仪比较BMDM的细胞形态及纯度;通过吞噬实验、杀菌实验以及液相芯片技术(liquid chip)比较两组BMDM的生物学功能;同时将上述结果与体内分化成熟的腹腔巨噬细胞相比较。[结果]两组诱导分化成熟的BMDM均呈现不规则形或梭形;BMDM的纯度分别为M-CSF组98.6%,L929上清组99.6%;腹腔细胞贴壁后巨噬细胞的纯度是98.8%;吞噬实验、杀菌实验两组之间及其与腹腔巨噬细胞相比无显著差异;炎症因子释放水平检测中3组细胞存在IL-6或TNF-α某些时间点释放水平差异。[结论]M-CS与L929上清诱导方式所获得BMDM的形态相似,纯度相近(98.6%比99.6%),吞噬细菌能力无显著统计学差异(21.31±5.83比26.10±6.11,t=-0.745,P>0.05),杀菌能力无显著差异(杀菌30min:36.41%±4.21%比39.53%±6.75%;60min:44.35%±6.75%比45.27%±1.96%,两者均P>0.05),但是炎症因子释放的某些时间点有差异(TNF-α的释放在LPS刺激后24 h M-CSF组高于L929组:549.92±412比271.47±432,t=-0.34,P<0.05),应根据实验目的选择获取BMDM的诱导方式。
[Objective] The aim of the study was to provide morphological and basic biological functions of bone marrow macrophage (BMDM) by comparing two commonly used M-CSF and L929 cell culture supernatants. [Method] The bone marrow of mice was induced by two ways to obtain BMDM. Then the cell morphology and purity of BMDM were compared by Zeiss fluorescence microscope and flow cytometry. By phagocytosis test, bactericidal test and liquid chip comparison Group BMDM biological function; at the same time, the above results and in vivo differentiation of mature peritoneal macrophages compared. [Results] The BMDM induced by both groups showed irregular or spindle shape. The purity of BMDM was 98.6% in M-CSF group and 99.6% in L929 supernatant group, respectively. The purity of peritoneal macrophages after adherence was 98.8 %; Phagocytosis test, bactericidal test between the two groups and compared with the peritoneal macrophages no significant difference; the release of inflammatory cytokines in the three groups of cells IL-6 or TNF-α released at some point in time difference. [Conclusion] The morphology of BMDM obtained by M-CS and L929 supernatants was similar (98.6% vs. 99.6%). The phagocytosis of bacteria was not statistically significant (21.31 ± 5.83 vs. 26.10 ± 6.11, t = -0.745 , P> 0.05). There was no significant difference in the bactericidal activity (bactericidal 30min: 36.41% ± 4.21% vs 39.53% ± 6.75%; 60min: 44.35% ± 6.75% vs 45.27% ± 1.96%, both P> 0.05) The release of inflammatory cytokines differed at some time points (TNF-α release was higher in the M-CSF group than in the L929 group at 24 h after LPS stimulation: 549.92 ± 412 vs. 271.47 ± 432, t = -0.34, P <0.05) According to the purpose of experiment choose to get BMDM induction.