直肠癌患者外周血单核细胞诱导为树突细胞

Research on peripheral blood mononuclear cells in patients with rectal cancer induced into dendritic cells

目的用直肠癌患者外周血单核细胞通过体外培养的方法诱导为成熟树突细胞。方法 CS-3000血细胞分离机采集直肠癌患者外周血单核细胞悬液,Ficoll分离单个核细胞,用含10%FBS的RPMI-1640培养基培养,第0天加入GM-CSF1000 U/ml、IL-4 500 U/ml,第4天2/3量换液,补足GM-CSF、白介素(IL)-4,并加入肿瘤坏死因子(TNF)-α500 U/m l诱导其成熟,以后每隔3天2/3量换液。结果培养至第11天,收集培养的1×105～2×105个树突细胞经流式细胞仪检测表达CD83+(52.9±5.8)%,CD86+(79.2±8.1)%,HLA-DR(65.8±5.7)%,CD1a(51.3±6.4)%,及CD14+(4.72±2.16)%。结论直肠癌患者外周血单核细胞经GM-CSF、IL-4及TNF-α联合培养能诱导出成熟树突细胞。为直肠癌的免疫治疗提供了临床应用基础。

Objective To induce the peripheral blood mononuclear cells（PBMCs） from rectal cancer patients into dendritic cells in vitro.Methods PBMCs of patients with rectal cancer were collected by cell detachment machine,abstracted by Ficoll,cultured in RPMI-1640 nutritive medium containing 10% FBS,induced by adding GM-CSF 1 000 U/ml,IL-4 500 U/ml on the 1st day,and 2/3 volume of fluid was exchanged,GM-CSF and IL-4 were complimented and TNF-α 500 U/ml was joined to promote mature on the 4th day,and thereafter 2/3 volume of exchange fluid every 3 days.Results 1×105~2×105 dendritic cells were collected and detected.The expression of CD83＋ was（52.9±5.8）%,CD86＋ was（79.2±8.1）%,HLA-DR was（65.8±5.7）%,CD1a was（51.3±6.4）%,and CD14＋ was（4.72±2.16）%.Conclusions PBMCs from rectal cancer patients can induced into dendritic cells by GM-CSF,IL-4 and TNF-α,and which provide the clinical application basis for cell immunotherapy of rectal cancer.