摘要
目的 探讨脐血树突状细胞 (DC)的体外诱导及其免疫学特征。方法 无菌条件下常规采集脐血 ,采用淋巴细胞分离液分离脐血单个核细胞 ,去除悬浮细胞后获得单核细胞。将获得的单核细胞分为两组 ,组 1:在含白介素 4(IL 4 )和粒细胞巨细胞集落刺激因子 (GM CSF)的DMEM液中培养。组 2 :在上述培养液中孵育 5天后 ,加入肿瘤坏死因子 (TNF α)。收集培养 7天的贴壁细胞 ,分别用流式细胞仪检测CD83、CD86 、CD54、CD1α和CD1 4等免疫因子。结果 ①组 1培养 12～ 14天树突状细胞数含量约 2 0 %。组 2培养 7～ 10天即可见到明显树突状细胞达 30 %。②TNF α可明显增加干细胞早期增殖能力 ,两组相比P <0 0 5。③IL 4、GM CSF和TNF α可诱导脐血来源的单核细胞 (即DC前体细胞 )分化为成熟的DC ,两组均高表达DC特异性抗原CD83、CD86 、CD54、CD1α。结论 脐血中的DC前体细胞可在体外诱导成为功能性 (即成熟 )DC。GM CSF联合TNF α不仅可以促进细胞形态的生长 ,而且明显增加DC生成数量。故脐血有可能成为DC免疫治疗丰富的来源。
Objective To investigate how to inducing dendritic cell(DC) from the precursor in cord blood and the immunological characteristics. Methods Cord blood was collected under steril condition and the mononuclear cells(MNCs) were separated by centrifugal in density gradient. The adhesion cells, collected among the MNCs, were divided into two groups. Group1:The adhesion cells were cultured with interleukin-4, granulocyte-macrophage colony-stimulating factor(GM-CSF) in dulbecco's modified eagles medium(DMEM) medium. Group2:Cultured in the above medium for the first 5 days, tumor necrosis factor α(TNF-α) was added subsequently. 7 days later, CD 83、CD 86、CD 54、CD 1α and CD 14 cell-surface phenotyping were examined by cytofluorimetry, respectively. Results ① Group1examined under phase microscopy revealed that the content of DC with the typical morphological characteristics increased by 12-14 day(the proportion to 20%), for Group2 it was 30% on day 7-10. ② TNF-α was demonstrated to be stronger stimulator for early proliferation of stem cell than GM-CSF(P<0.05). ③ Our results indicated that DC precursors in cord blood can be induced to mature DC in the medium containing IL-4, GM-CSF and TNF-α. High DC specialized antigen expression, such as CD 83、CD 86、CD 54 and CD 1α were seen. Conclusion Primary in vitro experiment has shown that the precursors in cord blood can be induced to fuctional DC. GM-CSF combined with TNF-α not only promote the development of cell morphology, but also increase obviously the amount of DC. Cord blood may become valuable origin for DC immunotherapy.
出处
《中国实验诊断学》
2004年第3期260-262,共3页
Chinese Journal of Laboratory Diagnosis
基金
国家自然科学基金资助项目 (编号 3 9970 770)
关键词
脐血
树突状细胞
粒细胞巨细胞集落刺激因子
肿瘤坏死因子
Cord blood
Dendritic cell
Tumor necrosis factor
Granulocyte-macrophage colony-stimulating factor
