【摘要】 目的研究多发性骨髓瘤(MM)患者外周血树突状细胞(DC)的分离、培养和鉴定。方法用塑料贴壁的外周血单个核细胞(PBMNC)来源或者免疫磁珠阴性选择分离途径,建立体外培养技术,从MM患者外周血中获取DC。结果(1)在含粒-巨噬细胞集落刺激因子和白细胞介素-4的无血清培养基(AIM-V)中,37℃、体积分数为0.05的CO2培养箱中培养5~7 d,可获得具有典型DC形态学特征的细胞。2种分离途径得率比较无统计学意义(n=5,P>0.05)。(2)同种混合淋巴细胞反应及自体抗原呈递试验证实,MM患者外周血来源DC具有很强的刺激同种异体淋巴细胞增殖能力及呈递自体抗原的能力。外周血来源DC培养5~7 d后流式细胞仪分析表明,部分细胞表达成熟DC特征性标记CD1a,部分表达共刺激分子CD80及CD86,高表达HLA-DR分子,几乎不表达单核细胞特异性标记CD14。重组人白细胞介素-3或/及重组人干细胞因子加入培养体系并不增加MM患者外周血DC得率。免疫表型也无明显改变。提示DC来源于PBMNC中单核细胞,而非CD34+造血祖细胞。结论贴壁分离法及免疫磁珠法均可获取充足数量的DC。更多还原

【Abstract】 Objective To set up an in vitro culture procedure obtaining derdritic cells（DC） from peripheral blood of multiple myeloma（MM） patients.Methods Plastic-adherent peripheral blood mononuclear cells（PBMNC） or sorted cells by immmunomagenetic separation method were cultured in AIM-V serum-free medium containing GM-CSF and IL-4 at 37 ℃,5%CO₂ humidified atmosphere.After 5～7 days of culture,a large quantities of cells exhibiting a typical characteristic dentritic cell morphology were collected.Results No significant difference in yield existed between two separation methods（n）.MLR assay and autologous antigen presentation assay showed that DC had potent capability to stimulate allogeneic lymphocyte proliferation.Cytometer analysis showed that some of cells expressed the characteristic marker CDla of mature DC.Costimulating molecules（CD80 and CD86）,HLA-DR molecules were highly expressed on DC.DC didn’t express CD14 monocyte marker although adding GM-CSF or/and IL-3 into culture medium.The immunophenotype of DC didn’t change that indicating that DC derived from monocyte,not from CD34⁺ progenitor cells.（Conclusion） Generation of DC from peripheral blood in MM patients may faciliate the further studies on DC and their clinical applications.更多还原