摘要
目的 研究人脐血来源的树突状细胞 (DCs)体外培养扩增 ,并检测其功能。方法 应用Ficoll Hypaque离心获得界面细胞 ,贴壁培养 2h ,获得单个核细胞 ,体外以重组hGM CSF( 5 0ng/ml) +hIL 4( 10ng/ml) +hTNF α( 5 0ng/ml)诱导培养 14天。在DCs发育过程中 ,在光镜下观察其生长情况 ,应用流式细胞仪检测DCs表型。采用MTT法检测DCs活化的T细胞对肿瘤细胞的杀伤性。结果 第 6天体外培养的DCs由贴壁状态变为悬浮毛刺状细胞 ,随着培养时间的延长 ,此类细胞数量增多 ,第 12天为形态不规则的毛刺状 ,为典型树突状细胞形态。流式细胞仪检测表明 ,成熟DCs高水平地表达HLA DR、CD 80、CD83、CD 1a、CD 11c、CD12 3。被激活的T细胞对 2种来源于不同组织的肿瘤细胞均产生了明显的杀伤性。结论 人脐血经rhGM CSF +rhIL 4+hTNF α体外诱导培养 ,能诱导出DCs。
Objective To investigate the proliferation and functions of dendritic cells derived from human cord blood progenitor cells in vitro.Methods The mononuclear cells were prepared from cord blood with Ficoll-hypaque centrifugation method,then induced with the recombinant cytokines hGM-CSF(50 ng/ml),hIL-4(10 ng/ml) and hTNF-α(50 ng/ml) for two weeks.We observed the DCs growth under the microscope and determined DCs phenotypes by flow cytometry.The capacity of DCs to activate T cell-depent anti-tumor immune responses was tested by MTT.Results The dendritic cells cultured in vitro turned into suspensive growth from adhesive situation at 6 d.Then the number of DCs increased continuously and the cells showed the irregular morphologic appearance of DCs with veiled edges on the 12th day.It was showed by flowcytometry that the mature DCs expressed the relatively specific marker such as HLA-DR,CD80,CD83,CD1a,CD11c and CD123.T cells activated by DCs showed magnificent cytotoxicity on two different kinds of tumor cells.Conclusion The dendritic cells can be induced from human cord blood through the cultivation with rhGM-CSF,rhIL-4 and rhTNF-α in vitro.
出处
《实用癌症杂志》
2004年第2期117-120,共4页
The Practical Journal of Cancer
