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小鼠骨髓树突状细胞的扩增及其超微结构观察
郑尊,于秉学,颜永碧,章卫平,曹雪涛,谢志芳
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摘要:
目的:建立体外大量扩增树突状细胞(DC)的方法,并对DC进行形态学和免疫学性质鉴定。方法:制备小鼠骨髓细胞,用免疫磁珠方法去除其中的淋巴细胞、单核巨噬细胞及粒细胞,将阴性选择到的细胞用小鼠重组GM-CSF培养,8 d后收集悬浮细胞。将收集到的细胞进行免疫表型鉴定和超微结构观察,以及DC特异性抗原DEC205的免疫胶体金标记。结果:可从每只小鼠的股骨骨髓中扩增到(2-5)×106 个DC, 细胞纯度达80%以上。细胞表达DC的特异性抗原DEC205,具有DC的典型形态及表型。结论:成功地建立了体外大量扩增骨髓DC的方法,为深入研究其免疫学功能打下基础。
关键词:  树突状细胞  超微结构
DOI:
基金项目:
Amplification and ultrastructure observation of murine bone marrow-derived dendritic cells
郑尊,于秉学,颜永碧,章卫平,曹雪涛,谢志芳
()
Abstract:
Objective: To amplify large number of dendritic cells (DCs) from murine bone marrow and assay their morphological, immunological characteristics. Methods: Lymphocytes, monocytes-macrophages and granulocytes of murine bone marrow were depleted using immuno-Minimagnetic cell sorter. The selected cells were cultured with granulocyte-macrophage colony-stimulating factor, and the floating cells were collected 8 d after culture. The phenotype of collected DCs was analyzed, and their morphology was observed under electron microscopy. Results: (2-5)×106 DCs were obtained from bone marrow of one mice, and the purity of DCs was more than 80%. The cells expressed specific antigens (DEC205) of DCs and had typical morphology of DCs. Conclusion: A method to generate large number of DCs from murine bone marrow is established, which contribute to further studying immunological functions of DCs.
Key words:  dendritic cells  ultrastructure