首都医科大学学报 ›› 2015, Vol. 36 ›› Issue (4): 610-613.doi: 10.3969/j.issn.1006-7795.2015.04.019

• 基础研究 • 上一篇    下一篇

一种简单的分离、培养及鉴定小鼠外周血单核巨噬细胞方法的建立

杨琳, 田蕾, 谢杰施, 李丽英   

  1. 首都医科大学细胞生物学系, 肝脏保护与再生调节北京市重点实验室, 北京 100069
  • 收稿日期:2015-02-08 出版日期:2015-08-21 发布日期:2015-07-17
  • 通讯作者: 李丽英 E-mail:liliying@ccmu.edu.cn
  • 基金资助:

    国家自然科学基金(81430013),北京市属高等学校创新团队建设与教师职业发展计划项目(IDHT20150502)。

Establishment of a simple method for separation, culture and identification of mouse peripheral blood monocyte/macrophage

Yang Lin, Tian Lei, Xie Jieshi, Li Liying   

  1. Department of Cell Biology, Municipal Laboratory for Liver Protection and Regulation of Regeneration, Capital Medical University, Beijing 100069, China
  • Received:2015-02-08 Online:2015-08-21 Published:2015-07-17
  • Supported by:

    This study was supported by National Natural Science Foundation of China(81430013), Project of Construction of Innovative Teams and Teacher Career Development for Universities and Colleges Under Beijing Municipality(IDHT20150502).

摘要:

目的 在L929条件培养基的作用下体外分离、培养小鼠外周血单核巨噬细胞,并进行鉴定。方法 密度梯度离心法分离小鼠外周血单核巨噬细胞,用L929条件培养基培养7 d。采用免疫荧光的方法检测F4/80的表达以及细胞的吞噬作用,用流式细胞术进一步检测细胞吞噬率。结果 用L929条件培养基诱导培养7 d后小鼠外周血单核细胞表达F4/80,加入细胞吞噬珠之后,在不同时间点用荧光显微镜可以看到红色荧光颗粒聚集在小鼠外周血单核巨噬细胞核周围,用流式细胞仪检测细胞的吞噬率为24.8%±0.79%。结论 该方法是一种简单、易操作的体外分离培养和鉴定小鼠外周血单核巨噬细胞的方法。

关键词: 小鼠, 外周血单核巨噬细胞, 分离培养, 鉴定

Abstract:

Objective The aim of this study is to separate, cultivate and identify mouse peripheral blood monocyte/macrophages(PBM) in L929-conditioned medium. Methods Mouse PBM was isolated by centrifugation, and cultured with L929-conditioned medium for 7 days. PBM was identified by immunofluorescence and flow cytometry(FCM). Results PBM display the immunophenotype of monocyte/macrophage and with phagocytic function. Conclusion It was a simple method for separation, culture and identification of mouse PBM in vitro.

Key words: mouse, peripheral blood macrophages, isolation and cultivation, identification

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