摘要
目的 :以大鼠骨髓造血干细胞为来源 ,建立体外诱导、扩增树突状细胞 (dendriticcells,DC)的方法 ,并进行表型鉴定及功能检测。 方法 :从大鼠骨髓细胞中分离出造血干细胞 ,加入rGM CSF、rIL 4和nrhTNF α培养 1 2天获得大量DC ,经标抗大鼠OX6 2单抗免疫磁珠分离纯化后的DC ,进行形态学观察、表型检测及刺激T细胞增殖能力分析。 结果 :大鼠骨髓造血干细胞经诱导培养 1 2天后 ,倒置显微镜和电镜下显示典型的DC形态和超微结构 ;流式细胞术分析显示细胞表面抗原高表达 ,其中OX6 2为 6 2 .1 9% ;MHCⅠ为 70 .4 0 % ;MHCⅡ为 78.2 8% ;CD80为 5 5 .5 8% ;CD86为 6 8.38% ,是典型的大鼠DC表型特征 ,同种混合淋巴细胞反应结果显示 ,DC具有高效的刺激T淋巴细胞活化增殖的能力。 结论 :大鼠骨髓造血干细胞可成功诱导为具有典型形态特征及功能的DC ,为进一步研究其在肿瘤免疫治疗中的应用打下基础。
Objective:To establish a method of inducing dendritic cells(DC)from the hematopoitic stem cells of rats in vitro,and to identify the phenotype and fusion of DC. Methods:DC obtained from SD rat bone marrow hematopoitic stem cells were propagated in vitro under the condition of rGM CSF、rIL 4 and nrhTNF α.DC were purified by monoclonal antibody OX62 and magnetic beads.Then DC harvested 12 d later were identified by morphological features,surface antigen expressions and the ability to atimulate T cells. Results:After culture and induction,DC displayed typical morphology with elongated dendritic processes viewed by inversion microscope as well as electron microscope.DC expressed high level surface antigens,including OX62 62.19%;MHCⅠ 70.40%;MHCⅡ 78.28%;CD80 55.58%; CD86 68.38%, The results of mixed lymphocyte reaction(MLR)showed that DC had the ability to stimulate vigorous proliferation of allo T cells. Conclusion:Matue DC could be generated from rat bone marrow hematopoitic stem cells,which presents the feasibility for further clinical application of DC in the immunotherapy of cancers.
出处
《医学研究生学报》
CAS
2004年第7期588-591,共4页
Journal of Medical Postgraduates
基金
国家自然科学基金资助项目 (批准号 :3 0 3 3 0 610 )
