抗CD11c磁珠阳性选择法分离肺癌组织浸润树突状细胞

Positive selection using anti-CD11c magnetic beads for isolation of tumor infiltrating dendritic cells in lung cancer tissues

目的：建立抗CDI1c磁珠阳性选择法分离Lewis肺癌模型中肿瘤浸润树突状细胞（tumor infiltrating dendritic cell，TIDC）的方法。方法：在C57BL／6小鼠侧腹壁皮下注射Lewis肺癌细胞（1．0×10^6／只）建立荷瘤小鼠模型。抗CD11c磁珠阳性选择法分离提取TIDC，抗小鼠CD11c-PE标记TIDC，用流式细胞仪检测分离到的细胞纯度；电镜观察细胞形态；抗小鼠MHC-Ⅱ-PE和CD83-FITC或CD86-FITC抗体双标记后，用流式细胞术检测细胞表型。结果：抗CD11c磁珠阳性选择法可在每克Lewis肺癌移植瘤组织中分离到（1．73±0．31）×10^6个TIDC，占肿瘤组织细胞总数的（2．18±0．29）％；TIDC纯度达到96．49％。电镜观察到分离的TIDC具有DC细胞的典型形态特征，其细胞表面MHC-Ⅱ、CD83和CD86分子的表达率分别为（51．25±4．21）％、（3．48±0．34）％和（3．07±0．65）％。结论：抗CD11c磁珠阳性选择法体外分离TIDC具有高效、简便、相对经济实用的优点，有推广价值。

Objective：To establish a method for isolating tumor infiltrating dendritic cells （TIDC） from mice models of Lewis lung cancer by positive selection using anti-CD11 c magnetic beads. Methods： A total of 1.0 × 10^6 Lewis lung cancer cells were subcutaneously injected into a C57BL/6 mice at the lateral abdominal wall to establish the mouse model of lung cancer. TIDC were isolated positively using anti-mouse CD11 c magnetic beads; they were labeled by anti -mouse CD11 c, and then the purity of the isolated cells was tested by FACScan flow cytometer. The cells were also double labeled by PE-conjugated MHC- Ⅱ/ mAb and FITC-conjugated CD83 mAb or FITC-conjugated CD86 mAb to analyze the phenotype of cells by FACScan flow cytometer. Results： The positive selection using anti-CD11c magnetic beads isolated （1.73± 0. 31 ） ×10^6 TIDC from each gram of Lewis lung cancer tissue, which accounted for （2.18 ±0.29）% of the cells in the tumor tissues. The purity of TIDC was 96.49%. Electron microscope showed that the isolated TIDC had the typical character of DC cells. The positive rates of MHC-Ⅱ , CD83 and CD86 molecules in TIDC surface were （51.25 ±4.21 ）%, （3.48 ± 0.34） % and （ 3.07 ± 0.65 ） %, respectively. Conclusion ： The positive selection using anti-CD11 c magnetic beads is highly effective, simple, and economic, and is worth popularizing.