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目的:探索树突状细胞(DC)及其前体的分离纯化及其体外扩增的方法。方法:无菌制备BALB/C小鼠骨髓和脾细胞;先后用红细胞裂解液、抗鼠CD4、CD8、B细胞单抗(McAb)和补体溶液,依次去除红细胞,T、B细胞,粒细胞和单核巨噬细胞等混杂细胞而获得纯化的DC及其前体;又在粒细胞巨噬细胞集落刺激因子(GMCSF)和白细胞介素(IL4)协同诱导下培育,DC前体分化发育成DC或郎罕细胞(LC)并扩增,同时阻抑巨噬细胞发育生长。结果:DC/LC细胞数增加,其形态在光镜下多为特征性星形,也有梭形和多角形;扫描电镜下观察其绝大多数为特征性星形,且有1~4级不等的树突状突起,其纯度高达95%以上。DC/LC在功能上明显刺激同种异体混合淋巴细胞反应(MLR)。结论:①结果所得细胞的形态和功能符合DC/LC;②建立连续排异结合GMCSF+IL4联合诱生培育的方法,可获得大量高纯度的DC/LC。
Objective: To explore the isolation and purification of dendritic cells (DCs) and their precursors and their in vitro amplification methods. Methods: BALB / C mouse bone marrow and spleen cells were prepared aseptically. Erythrocyte, T, B cells, granulocytes and Mononuclear macrophages and other hybrid cells to obtain purified DC and its precursor; and granulocyte-macrophage colony stimulating factor (GM CSF) and interleukin (IL 4) under the co-induced cultivation, DC Precursors differentiate into DCs or Langerhans cells (LCs) and proliferate, inhibiting the growth of macrophages. Results: The number of DC / LC cells increased. The morphology of the DC / LC cells was mostly characterized by light microscopy with spindles and polygons. Most of them were characterized by scanning electron microscopy, Dendritic protuberances, the purity of up to 95%. DC / LC functionally stimulates allogeneic mixed lymphocyte reaction (MLR). CONCLUSION: ① The morphology and function of the obtained cells are in accordance with DC / LC. ②A series of high-purity DC / LC can be obtained by establishing a method of continuous exclusion combined with GM-CSF + IL-4 induction.