小鼠骨髓源性树突状细胞的体外诱导扩增和鉴定

发布时间：2018-02-13 17:49

  本文关键词： 树突状细胞 诱导 鉴定　出处：《华中科技大学学报(医学版)》2013年04期 　论文类型：期刊论文

【摘要】：目的体外诱导扩增获取大量高纯度的小鼠骨髓来源树突状细胞(DC),并研究不同生长状态的DC成熟情况及进行生物学特性鉴定。方法以重组小鼠粒细胞巨噬细胞集落刺激因子(rmGM-CSF)和重组小鼠白细胞介素-4(rmIL-4),体外诱导小鼠骨髓细胞分化为DC,脂多糖(LPS)刺激24h,倒置显微镜动态观察细胞形态学变化,并分别收集悬浮细胞、贴壁细胞,流式细胞术(FACS)分析DC细胞表面分子CD11c、MHCⅡ类分子、CD80和CD86的表达水平,并应用混合淋巴细胞反应(MLR)检测其刺激异基因淋巴细胞增殖的能力。结果经体外诱导培养第3天即可见大量细胞集落形成;培养至第9天,细胞形态不规则,具有典型DC形态。同组悬浮DC表面MHCⅡ类分子和共刺激分子CD80、CD86的表达水平高于贴壁DC。LPS刺激24h后,DC表面MHCⅡ类分子、CD80、CD86的表达水平显著升高,同时可以显著刺激同种异基因淋巴细胞增殖。结论体外简易诱导培养可以获得高纯度小鼠骨髓来源的各具特性的DC,为开展DC的相关实验研究提供大量稳定的细胞来源。
[Abstract]:Objective to obtain a large number of high purity mouse bone marrow derived dendritic cells (DC) by induction amplification in vitro, and to study the maturation and biological characteristics of DC in different growth states. Methods the colony of murine granulocyte macrophages was obtained by recombinant mouse granulocyte macrophage colony. The stimulating factor rmGM-CSF and recombinant mouse interleukin-4rmIL-4 were induced to differentiate into DCand lipopolysaccharide (LPS) for 24 h. The morphological changes were observed by inverted microscope. Suspension cells, adherent cells and FACS were collected to analyze the expression levels of CD11cMHC class 鈪，

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