【摘要】 目的　探讨从小鼠骨髓分离纯化及大量扩增树突细胞 (DC)的方法 ,并对其行免疫表型和形态鉴定。　方法　制备 Balb/ c小鼠骨髓细胞 ,利用 0 .83% NH4 Cl- Tris溶液、抗鼠 CD4 / CD8/ CD4 5R单克隆抗体和补体溶液 ,依次去除红细胞、淋巴细胞、单核 -巨噬细胞、粒细胞等混杂细胞以获得纯化的 DC,再将其培养于含有小鼠重组的粒细胞 -巨噬细胞集落刺激因子 (m GM- CSF)和白细胞介素 4 (m IL - 4 )的 RPMI 1 6 4 0营养液中 ,培养第 6～ 8天收集悬浮的细胞 ,电子显微镜观察细胞形态并用流式细胞术检测小鼠 DC细胞表面特有的 CD1 1 b抗原和 MHC- 类分子的表达量。　结果　每只小鼠可扩增到 (2～ 4 )× 1 0 6个 DC细胞 ,细胞纯度 >85 % ;细胞表面高水平表达小鼠 DC细胞特异性抗原 CD1 1 b和 MHC- 类分子 ,电镜下细胞形态符合典型的树突细胞形态。　结论　联合应用 m GM- CSF和m IL- 4刺激小鼠骨髓细胞可扩增到大量高纯度的树突细胞更多还原

【Abstract】 Objective To isolate and amplify large number of dendritic cells(DC) from mouse bone marrow and identify their morphological and immunological characteristics. Methods Bone marrow suspensions were prepared from Balb/c mouse, then the erythrocytes, lymphocytes, monocytes macrophages and granulocytes were removed by using 0 83%NH 4Cl Tris buffer, mouse anti CD 4/CD 8/CD 45R monoclonal antibodies and goat complement buffer. Selected DC cells were cultured in RPMI 1640 containing both recombinant mouse granulocyte macrophage colony stimulating factor(mGM CSF) and interleukin 4(mIL 4). The floating cells were collected after 6~8 days of culture. The immunological phenotype of collected DC was analysed by FACScan, and their morphology was observed under electron microscopy. Results (2~4)×10 6 dendritic cells were obtained from bone marrow of every mice, and the purity of DC was more than 85%. The DC were expressed with high level of CD 11b and MHC class Ⅱ molecules, which were specific antigens of mouse bone marrow derived dendritic cells. The collected cells had typical shape of DC also. Conclusion A large number of high purity dendritic cells could be amplified under the aegis of mGM CSF and mIL 4.更多还原