人外周血树突状细胞的分离培养及鉴定

目的从外周血中分离单个核细胞 ,经体外诱导培养获取高纯度的树突状细胞。方法淋巴细胞分离液分离外周血的单个核细胞 ,经贴壁 2h后加入GM CSF、IL 4诱导 7天 ,观察细胞形态 ,用流式细胞仪检测其表面标志 ,采用混合淋巴细胞培养法检测其刺激同种异体淋巴细胞增殖的能力。结果经诱导生成的细胞具有典型的DCs形态特征 ,高表达CD83 、CD86和MHCⅠ、Ⅱ类分子 ,在体外能诱导强烈的同种异体淋巴细胞增殖反应。结论本研究所建立的经济、简便的从外周血获取大量成熟DCs的方法 ,为DCs的进一步研究奠定了基础。

Identification and inculation of dendritic cells from human peripheral blood

objective To get high quality and sufficient numbers of mature dendritic cells (DCa) Methods Mononuclear cells was isolated from human peripheral blood by ficoll centrifugation and were adhered in plate. The adherent cells were then cultured with granulocyte/macrophage colony- stimulating factor (GM-CSF) and inteleukin 4(11-4). The cellular morphology was checked. The surface markers and the MLR stimulatory capacity of DCs were assayed by FACS and by 3H -TDR. Results Most suspended cells exhibite distinctive morphological features of DCs and they also can stimulate proliferation of allogeaic lymphcytes. Conclusion These results indicate that large numbers of DCs can be obtained from human peripheral blood and may facilitate further studies of DCs.