未成熟树突状细胞快速分离与体外诱导培养及其鉴定研究

发布时间：2018-06-09 05:01

本文选题：单核细胞 + 未成熟树突状细胞　；参考：《中国免疫学杂志》2017年07期

【摘要】：目的:以人外周血来源的单核细胞为前体细胞,建立体外快速分离和诱导培养未成熟树突状细胞(Immature dendritic cell,iDC)的方法。方法:采用Ficoll密度梯度离心方法和MACS磁珠分选系统,收集高纯度CD14~+单核细胞;用rhGM-CSF、rhIL-4联合诱导培养CD14~+单核细胞,第4天获得未成熟树突状细胞(iDC),应用流式细胞术检测细胞表面标记(CCR5)和抗原吞噬能力,普通光学显微镜、扫描电镜和透射电镜观察细胞表面和内部结构特征。结果:CD14免疫磁珠技术获得CD14~+单核细胞纯度达94%以上,诱导分化第4天的iDC具有CCR5特征性标志和吞噬能力,普通光学显微镜及电镜观察到iDC出现树突状细胞典型特征。结论:体外快速诱导培养是获得大量具有典型特征的iDC的有效方法,并可应用于进一步实验研究。
[Abstract]:Objective: to establish a method for rapid isolation and induction of immature dendritic cells (Immature dendritic cell, iDC) in vitro from monocyte derived from human peripheral blood. Methods: Ficoll density gradient centrifugation and MACS magnetic bead sorting system were used to collect high purity CD14~+ mononuclear cells, and rhGM-CSF and rhIL-4 were used to induce culture of CD14~+ mononuclear cells. CD14~+ mononuclear cells were cultured and immature dendritic cells (iDC) were obtained at fourth days. Flow cytometry was used to detect cell surface markers (CCR5) and antigen phagocytosis. Common optical microscopy, scanning electron microscopy and transmission electron microscopy were used to observe the surface and internal structure characteristics of the cells. Results: the purity of CD14~+ mononuclear cells obtained by CD14 immunomagnetic beads was more than 94%, IDC with fourth days of induced differentiation has CCR5 characteristic signs and phagocytosis. The typical characteristics of dendritic cells in iDC are observed by ordinary optical microscopy and electron microscopy. Conclusion: rapid induction and culture in vitro is an effective method for obtaining a large number of typical features of iDC and can be applied to a further experimental study.
【作者单位】：青岛市中心医院药学部;青岛市肿瘤医院药学部;军事医学科学院军事兽医研究所省部共建吉林省人兽共患病预防与控制重点实验室;长春中医药大学基础医学院;
【基金】：国家自然科学基金(31472197) 病原微生物生物安全国家重点实验室开放课题(SKLPBS1435) 北京市自然科学基金(5152023) 吉林省青年科研基金项目(20140520173JH)资助项目
【分类号】：R392

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