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目的探索从SD大鼠肝脏组织中直接提取树突状细胞(dendritic cell,DC)以供研究的方法。方法对SD大鼠肝脏先行二步灌流法灌流,再予密度梯度离心法离心,从而获得肝脏组织非实质细胞,接着应用大鼠树突状细胞特异性抗体OX62单抗分选出肝脏组织来源的树突状细胞,并且在光学显微镜及电子显微镜下观察其细胞形态,再结合流式细胞仪鉴定树突状细胞的表型。结果单只SD大鼠肝脏组织中可获得(0.7～1.0)×106个细胞,OX62单抗阳性率在60.92%～88.71%之间,顺利经过了其形态学及特异性表面标志角度的鉴定。结论初步建立了从SD大鼠直接分离提取肝脏组织树突状细胞的方法,为后期研究打下一定的基础。
Objective To explore a method for direct extraction of dendritic cells (DCs) from liver tissues of SD rats. Methods The SD rat liver was perfused by two-step perfusion and then centrifuged by density gradient centrifugation to obtain non-parenchymal cells in liver tissue. Then the rat dendritic cell-specific antibody OX62 monoclonal antibody was used to separate the liver tissue-derived Dendritic cells were observed under light microscope and electron microscope. The dendritic cells were identified by flow cytometry. Results The positive rate of OX62 monoclonal antibody in the livers of SD rats was (0.7-1.0) × 106 cells. The positive rate of OX62 was between 60.92% and 88.71%. The morphological and specific surface markers were successfully identified. Conclusion The method of direct separation and extraction of dendritic cells from the liver of SD rats was established initially, which laid the foundation for the later study.