【摘要】 目的　建立人脾脏体外培养树突状细胞的新方法。方法　人脾脏细胞悬液培养 2h后获得贴壁的单核细胞 ,加入重组人粒细胞巨噬细胞集落刺激因子 (rhGMCSF) 10 0 μg/L或rhGMCSF 10 0 μg/L +重组人白细胞介素 4 (rhIL 4 ) 5 0 0k/L ,体外培养 1周 ,收集悬浮细胞 ,以流式细胞仪分析细胞表型及抗原内吞能力 ,alloMLR检测细胞抗原呈递功能。结果　以细胞因子培养 7d生成的悬浮细胞 ,2 0 %～ 80 %表达树突状细胞特异性标志CD1a ,表达高水平的MHC Ⅱ、B71、B72分子 ,具有较强的抗原内吞能力 ,抗原内吞能力及T淋巴细胞活性与人外周血单核细胞培养获得的DC相似。结论　从人脾脏获得的贴壁的单核细胞 ,体外以rhGMCSF +rhIL 4培养 ,可以获得极其大量的树突状细胞更多还原

【Abstract】 Objective To explore the new method for the culture of human spleen dendritic cell(DC) Methods Human spleen cells were cultured for 2 hours at 37℃. The aderent cells were selected and then further cultured in medium with rhGM CSF(100μg/L)and rhIL4(500k U/L) for a week The endocytosis activity and T lymphocyte activity of aderent cell cultured were analysed with flow cytometry and allo MLR respectively Results DC specific marker(CD1a) were positive on 20%～80% of the cultured aderent cell High levels of MHC Ⅱ B71, B72 expression were also observed on the surface of the cell cultured The endocytosis activity and T lymphocyte activity of the cultured cell were similar to human spleen DCs and pheripheral moinocyte derived DCs Conclusion Large quantities of DC can be harvested by culturing the adherent human spleen monocytes with rhGM CSF and rhIL4 in vitro更多还原