Generation of large numbers of dendritic cells from mouse bone marrow cultures supplemented with granulocyte/macrophage colony-stimulating factor.
Generation of large numbers of dendritic cells from mouse bone marrow cultures supplemented with granulocyte/macrophage colony-stimulating factor.
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DOI：
10.1084/jem.176.6.1693
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发表时间：
1992-12-01
期刊：
影响因子：
--
通讯作者：
Steinman RM
中科院分区：
文献类型：
--
作者：
Inaba K;Inaba M;Romani N;Aya H;Deguchi M;Ikehara S;Muramatsu S;Steinman RM
已知富含抗原呈递、主要组织相容性复合体（MHC）II类的树突状细胞来自骨髓。然而，骨髓缺乏成熟的树突状细胞，大量增殖的不太成熟的细胞还有待鉴定。最近描述的用于诱导小鼠血液中树突状细胞生长的方法现在已经被修改为骨髓中的MHC II类阴性前体。一个关键步骤是在培养的前2-4天通过温和洗涤去除大部分未粘附的新形成的粒细胞。这留下了松散地附着在更牢固的粘附“基质”上的增殖簇。“在第4-6天，这些细胞团可以被移走，通过1克沉淀分离，在重新培养时，释放出大量的树突状细胞。后者很容易根据其独特的细胞形状、超微结构和抗原库进行鉴定，如用一组单克隆抗体检测的那样。树突状细胞表达高水平的MHC II类产物，并作为启动混合白细胞反应的强大辅助细胞。如果应用巨噬细胞集落刺激因子而不是粒细胞/巨噬细胞集落刺激因子（GM-CSF），则既不产生簇也不产生成熟的树突状细胞。因此，GM-CSF产生所有三种髓样细胞谱系（粒细胞、巨噬细胞和树突细胞）。由于> 5x 10（6）树突状细胞在1周内从单个动物的大后肢骨内的前体发育而来，因此骨髓祖细胞可以作为树突状细胞的主要来源。这一特点应证明是有用的，否则这种微量细胞类型的未来分子和临床研究。
Antigen-presenting, major histocompatibility complex (MHC) class II- rich dendritic cells are known to arise from bone marrow. However, marrow lacks mature dendritic cells, and substantial numbers of proliferating less-mature cells have yet to be identified. The methodology for inducing dendritic cell growth that was recently described for mouse blood now has been modified to MHC class II- negative precursors in marrow. A key step is to remove the majority of nonadherent, newly formed granulocytes by gentle washes during the first 2-4 d of culture. This leaves behind proliferating clusters that are loosely attached to a more firmly adherent "stroma." At days 4-6 the clusters can be dislodged, isolated by 1-g sedimentation, and upon reculture, large numbers of dendritic cells are released. The latter are readily identified on the basis of their distinct cell shape, ultrastructure, and repertoire of antigens, as detected with a panel of monoclonal antibodies. The dendritic cells express high levels of MHC class II products and act as powerful accessory cells for initiating the mixed leukocyte reaction. Neither the clusters nor mature dendritic cells are generated if macrophage colony-stimulating factor rather than granulocyte/macrophage colony-stimulating factor (GM-CSF) is applied. Therefore, GM-CSF generates all three lineages of myeloid cells (granulocytes, macrophages, and dendritic cells). Since > 5 x 10(6) dendritic cells develop in 1 wk from precursors within the large hind limb bones of a single animal, marrow progenitors can act as a major source of dendritic cells. This feature should prove useful for future molecular and clinical studies of this otherwise trace cell type.