骨髓单个核细胞分化为CD103^+树突状细胞的方法研究（英文）  

Methods of CD103^+ dendritic cells differentiation from bone marrow

作　　者：陈云秀 吴毅 彭欣 孔潇 李媛媛 田小银 张光莉[3] 罗征秀[3] Yunxiu Chen;Yi Wu;Xin Peng;Xiao Kong;Yuanyuan Li;Xiaoyin Tian;Guangli Zhang;Zhengxiu Luo(Chongqing Three Gorges Central Hospital,Chongqing 404000,China;Key Laboratory of Child Development and Disorders of Ministry of Education,Chongqing 400014,China;Department of Respiratory Medicine,Children's Hospital of Chongqing Medical University,Chongqing 400014,China)

机构地区：[1]重庆市三峡中心医院江南分院,重庆404000 [2]儿童发育疾病研究省部共建教育部重点实验室,重庆400014 [3]重庆医科大学附属儿童医院呼吸科,重庆400014

出　　处：《广西医科大学学报》2019年第12期1882-1888,共7页Journal of Guangxi Medical University

基　　金：supported by the National Natural Science Foundation of China(No. 81470222);the Science and Technology Project of Chongqing(No. cstc2017jcyj B0160);the Science and Technology Project of Yuzhong District,Chongqing(No. 20170120)

摘　　要：目的:探索小鼠骨髓单个核细胞在体外培养诱导其分化为CD103^+树突状细胞(DCs)的方法,为免疫性疾病的防治提供实验基础。方法:分离小鼠胫腓骨,冲洗骨髓腔得到骨髓细胞,利用单个核细胞分离液分离出单个核细胞,加入粒细胞巨噬细胞集落刺激因子(GM-CSF)、Fms样酪氨酸激酶3配体(FLT3L)诱导其分化为成熟的DCs。显微镜下观察细胞大小、形态、分布,流式细胞术检测细胞CD103^+及其共刺激分子的表达水平,再利用磁珠分选出CD103^+DCs,检测其纯度。结果:单个核细胞培养8 d后,倒置显微镜下观察到细胞分化为多种形态,大部分呈圆形或不规则,大小不一,细胞周围有细小凸起,成簇状分布,细胞表面CD11c和CD103增多,表面标志物CD40、CD80、CD86和MHC-Ⅱ升高,磁珠分选纯化后CD103^+DCs纯度可达95.7%,分选后的CD103^+DCs可有效促进T细胞分化。结论:利用GM-CSF和FLT3L联合诱导法进行体外培养,能够得到纯度较高的CD103^+DCs。Objective:This study was performed to explore methods of inducing and culturing CD103^+dendritic cells(DCs)from mouse bone marrow mononuclear in vitro.Methods:The bone marrow mononuclear cells were obtained from femurs and tibias,and purified by mononuclear cells separation solution.Cells incubated with granulocyte-macrophage colony-stimulating factor(GM-CSF)and Fms-like tyrosine kinase-3 ligand(FLT3L)were added to induce cell differentiation.The morphology,phenotype and distribution were observed under invert microscope.The expression of CD103^+DCs and co-stimulatory molecules were detected by flow cytometry.CD103^+DCs were sorted by magnetic beads and detected the purity.Results:After 8 days culture,the mononuclear cells were differentiated into various forms under the microscope.Most of them were round or irregular,various morphologies and distribute clustered with plenty of dendrites.The expression of CD11c,CD103 and co-stimulatory molecules elevated significantly,and the purity of CD103^+ DCs was 95.7%.Besides,the CD103^+DCs could stimulate T cell differentiation efficiently.Conclusion:CD103^+DCs can be efficiently induced by GM-CSF and FLT3L in vitro.

关 键 词：单个核细胞 CD103^+ DCs GM-CSF FLT3L 

分 类 号：R392.3[医药卫生—免疫学]