论文部分内容阅读
目的 观察体外富集的人外周血树突状细胞（ｄｅｎｄｒｉｔｉｃ ｃｅｌｌｓ，ＤＣ） 的光镜形态学，细胞超微结构及细胞化学特征。方法 取健康人新鲜外周抗凝血，采用淋巴细胞分离液分离单个核细胞进行培养，经细胞因子（ｒｈＧＭＣＳＦ，ｒｈＩＬ４ ，ｒｈＴＮＦα） 联合刺激培养９～１２ 天，富集含ＤＣ的细胞群，光镜形态学分辨ＤＣ形态及含量，用抗人血ＤＣ表面特异单克隆抗体（ｍＡｂ Ⅻ） 确认ＤＣ的细胞化学特征。结果 体外培养富集的人外周血ＤＣ群的ＤＣ含量达７０ ％ ～８０ ％ 。透射电镜观察表明，细胞因子诱导的人外周血ＤＣ群可见到两类ＤＣ亚群：一类可能来自ＤＣ前体细胞，其细胞表面有大量不规则树枝状突起（ 粗细不等） ，细胞核不规则，核仁小，细胞化学染色［髓过氧化物酶（ＰＯＸ） 、非特异性酯酶（ＮＳＥ）］ 均为阴性；另一类是由单核细胞分化来的ＤＣ，细胞形态不规则，细胞表面粗糙，有大量皱褶及较粗大树枝状突起，胞核常呈现肾形或马蹄形，核内异染色质边集。结论 细胞因子诱导的人外周血ＤＣ群存在多相性。
Objective To observe the light microscopic morphology, cell ultrastructure and cytochemical characteristics of dendritic cells (DCs) enriched in vitro. Methods The fresh peripheral anticoagulant blood of healthy people was collected. Lymphocyte separation liquid was used to isolate mononuclear cells for culture. After cultured with rhGMCSF, rhIL4 and rhTNFα for 9 ~ 12 days, The morphological features of DCs were identified by light microscopy. The cytochemical characteristics of DCs were confirmed by anti-human DCs with surface-specific monoclonal antibodies (mAbs). Results The DC content of DCs cultured in vitro was 70% ~ 80%. Transmission electron microscopy showed that two types of DC subpopulations were found in cytokine-induced human peripheral blood DCs: one may come from DC precursor cells, the surface of which has a large number of irregular dendrites (ranging in thickness), irregular nuclei , Small nucleolus, cytochemical staining (myeloperoxidase (POX), non-specific esterase (NSE)] were negative; the other is from monocytes differentiated DC, irregular cell morphology, the cell surface Rough, a large number of folds and thicker dendrites, the nucleus often reniform or horseshoe-shaped nucleus heterochromatin edge set. Conclusion Cytokine induced human peripheral blood DC population heterogeneity.