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应用细胞分层剂密度梯度离心辅以细胞贴壁和花环沉降法,分离和纯化人外周血树突状细胞。通过Et花环形成细胞测定、荧光染色镜检和FACS分析表明,分离的细胞纯度可达40%~60%,残留的其他细胞多为NK细胞、单核细胞和T细胞。FACS分析还显示:所分离的细胞表面MHCⅡ类抗原的阳性率达97.34%;扫描电镜观察可见独特的细胞形态。将此类细胞加入体外淋巴细胞增殖反应体系则呈现明显促进作用。提示:分离的细胞形态与功能均与树突状细胞相符。
Human peripheral blood dendritic cells (DCs) were isolated and purified by density gradient centrifugation with cell decker and cell attachment and rosette sedimentation. The results of FACS analysis showed that the purity of isolated cells was up to 40% -60%. Most of the other remaining cells were NK cells, monocytes and T cells. FACS analysis also showed that the positive rate of MHCⅡ antigen isolated on the cell surface was 97.34%, and the unique cell morphology was observed by scanning electron microscopy. The addition of such cells to lymphocyte proliferation in vitro reaction system showed a significant role in promoting. Tip: The shape and function of isolated cells are consistent with dendritic cells.