乳腺癌患者外周血干细胞诱导为树突细胞的实验研究  

Peripheral blood mononuclear cells in patients with breast cancer induced by dendritic cells in vitro Treatment

作　　者：韩光宇[1,2] 任思坡[1,2] 谭昆[1,2] 拾莉[1,2] 耿跃春 梁军[1,2] 

机构地区：[1]徐州市医学科学研究所 [2]徐州市中心医院徐州市细胞生物治疗重点实验室,江苏徐州221006

出　　处：《中国实验诊断学》2013年第5期837-840,共4页Chinese Journal of Laboratory Diagnosis

基　　金：徐州市科技局社会发展项目XF10C087;XZZD1044;XZZD1045;徐州市科技局发展基金项目XM09B044

摘　　要：目的乳腺癌患者经动员后采集外周血干细胞,通过体外培养的方法诱导为成熟树突细胞。方法乳腺癌患者注射G-CSF进行干细胞动员3天,血细胞分离机采集外周血干细胞悬液,用Ficoll分离,收集单个核细胞,RP-MI-1640培养基重悬静置30min,贴壁细胞用含GM-CSF 1 000U/ml、IL-4 500U/ml、10%FBS的RPMI-1640培养基继续培养,第4天2/3量换液,补足GM-CSF、IL-4,并加入TNF-α500U/ml,第7天换液,补足细胞因子。结果培养至第9天,收集培养的1-2×105个树突细胞经流式细胞仪检测表达CD83+(47.8±6.2)%,CD86+(83.2±10.1)%,HLA-DR(75.9±7.9)%,CD1a(60.3±6.8)%。结论动员后的乳腺癌患者外周血干细胞经GM-CSF、IL-4及TNF-α联合培养能诱导出成熟树突细胞,为乳腺癌的免疫治疗提供了临床应用基础。Abstract：Objective The peripheral blood mononuelear cells from breast cancer patients induced into dendritic cells in vitro. Methods After stem cells mobilization the peripheral blood mononuelear cells （PBMC） of patients with breast cancer were collected by cell detachment machine,abstracted by Ficoll,suspended by RPMI 1640 nutritive medium and standing for 30 minute, then the adherent cells were cultured in RPMI-1640 nutritive medium containing 10 percent FBS, induced by adding GM-CSF 1000U/ml, IL-4 500U/ml, the 4th day 2/3 volume of exchange fluid, fill the GM-CSF, IL-4,and join the induction of TNF-α 500 U/ml mature,and the 7th day 2/3 volume of exchange fluid,making up for cytokine. Results On the 9th day we collected 1 2×10^5 dendritic cells and detected CD83＋ （47.8±6.2）%, CD86 （83.2±10.1）,HLADR （75.9±7.9）%,CDla（60.3±6.8）%.Conelusion After stem cells mobilization the PBMC from breast cancer patients can be induced into dendritic cells by adding GM-CSF, IL 4 and TNF-a, and will provide the basis of clinical application for cell immunotherapy of breast cancer patients.

关 键 词：乳腺癌 外周血干细胞 树突细胞 免疫治疗 

分 类 号：R737.9[医药卫生—肿瘤]