小鼠骨髓树突状细胞的扩增及其超微结构观察

Amplification and ultrastructure observation of murine bone marrow derived dendritic cells

目的：建立体外大量扩增树突状细胞（ Ｄ Ｃ）的方法，并对 Ｄ Ｃ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 Ｇ Ｍ Ｃ Ｓ Ｆ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 Ｄ Ｃ特异性抗原 Ｄ Ｅ Ｃ２０５ 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 Ｄ Ｃ， 细胞纯度达８０％ 以上。细胞表达 Ｄ Ｃ的特异性抗原 Ｄ Ｅ Ｃ２０５，具有 Ｄ Ｃ的典型形态及表型。结论：成功地建立了体外大量扩增骨髓 Ｄ Ｃ的方法，为深入研究其免疫学功能打下基础。

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)×10 6 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.