摘要
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.
出处
《中国肿瘤生物治疗杂志》
CAS
CSCD
2008年第6期581-584,共4页
Chinese Journal of Cancer Biotherapy
基金
复旦大学附属华山医院科研基金(No.145)~~