人脐血树突状细胞体外扩增的研究

Generation of dendritic cells from human cord blood

目的 探讨人脐血来源的树突状细胞 (dendriticcells,DC )的体外培养、增殖情况。方法 采用密度梯度离心法获得界面细胞 ,贴壁培养 2小时 ,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 2周。在DC发育过程中 ,在光镜下观察其生长情况 ,流式细胞仪检测DC表型。结果 体外培养的DC在第四天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 ,第八天为形态不规则的毛刺状 ,为典型树突状细胞形态。流式细胞仪显示体外培养成熟的DC高表达共刺激分子CD80、黏附分子CD5 4、MHCII类分子HLA DR。结论 人脐血经hGM CSF +hIL 4 +hTNF α体外培养 ,能诱导出DC ,本研究为DC的深入研究与临床应用奠定了基础。

Objective: To investigate the culture and proliferation of dendritic cells derived from cord blood 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 and determined DCs phenotypes by flow cytometry. Results: The dendritic cells culturedin vitro turned into suspensive growth from adhesive situation at 4d, along with the incubation, the number of DCs increased and the cells showed the irregular morphologic appearance of DCs with veiled edges in the 8th day. The mature DCs could express the relatively specific marker such as co-stimulatory molecules CD80, adhesion molecule CD54 and MHC-Ⅱmolecule HLA-DR. Conclusion: The dendritic cells can be induced from human cord blood that is co-incubated with hGM-CSF, hIL-4 and hTNF-α in vitro. The investigation establishes a foundation for furthur research and clinical use of DCs.