中国畜牧兽医

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3种分离小鼠腹腔肥大细胞方法的研究

谷玉静1，郝满良1，尹丽华2，董国权3   

  1. （1.河北农业大学动物医学院，河北保定 071001；2.邢台市桥西区农业局疫控中心，河北邢台 054000；3.辛集市畜牧局防疫监督站，河北辛集 052360）
  • 收稿日期:2013-07-16 出版日期:2014-02-20 发布日期:2014-03-27
  • 通讯作者: 郝满良（1959—），男，河北人，副教授，研究方向：肥大细胞分子免疫学。E-mail: haomanliang59@sohu.com
  • 作者简介:谷玉静（1987—），女，河北人，硕士，研究方向：天然免疫与动物疾病防治。
  • 基金资助:

    河北省自然科学基金（C2011204008）。

Study on Three Separation Methods of Mast Cells of Mice Abdominal Cavity

GU Yu-jing1, HAO Man-liang1, YIN Li-hua2, DONG Guo-quan3   

  1.  (1.College of Veterinary Medicine,Agricultural University of Hebei,Baoding 071001,China;2.Agricultural Bureau of Xingtai Qiaoxi District, Xingtai 054000,China;3.Xinji City Animal Husbandry Bureau,Xinji 052360, China)
  • Received:2013-07-16 Online:2014-02-20 Published:2014-03-27

摘要： 为获取小鼠腹腔肥大细胞，试验采用3种Percoll梯度分离法对小鼠腹腔肥大细胞进行了分离，对其细胞活率、细胞纯度及细胞总数进行了统计；并将离心管中3 mL Percoll梯度分离液自上而下分为4层来分析肥大细胞的动态分布。结果显示，方法Ⅰ分离到的腹腔肥大细胞活率为（83.51±14.00）%，纯度为（69.04±11.75）%，细胞总数为（10.60±5.20）×105 /mL；方法Ⅱ活率为（85.71±9.23）%，纯度为（87.10±3.93）%，细胞总数为（11.64±5.73）×105 /mL；方法Ⅲ活率为（72.25±24.81）%，纯度为（68.34±10.20）%，细胞总数为（8.87±5.18）×105/mL。其中活率、细胞总数在3种方法间差异均不显著（P＞0.05）；而方法Ⅱ的细胞纯度极显著高于方法Ⅰ和Ⅲ（P＜0.01）。肥大细胞在Percoll梯度分离液中的动态分布，3、4层的细胞活率均极显著高于1层（P＜0.01），均显著高于2层（P＜0.05）；3层肥大细胞纯度极显著高于1、4层（P＜0.01），显著高于2层（P＜0.05）；3、4层的细胞总数均极显著高于1、2层（P＜0.01）。因此，采用方法Ⅱ及第3层，即抽取中下0.5 mL，也就是接近30%∶80%的Percoll梯度分离液界面部位，分离获取小鼠腹腔肥大细胞效果最好。

关键词: 小鼠; 肥大细胞; 分离方法

Abstract: In order to get mast cells in mice abdominal cavity,we used three Percoll gradient separation methods to extract mast cells from mice abdominal cavity,and counted cell viability, cell purity and the total number of cells.The 3 mL Percoll gradient separation liquids in the centrifugal tube from top to bottom were divided into 4 layers to analyze the dynamic distribution of mast cells.The results showed that in the method Ⅰ,the cell viability, cell purity and the total number of cells were (83.51±14.00)%，(69.04±11.75)% and（10.60±5.20）×105 /mL,respectively；in the method Ⅱ,the cell viability, cell purity and the total number of cells were (85.71±9.23)%，(87.10±3.93)% and（11.64±5.73）×105 /mL,respectively；in the method Ⅲ,the cell viability, cell purity and the total number of cells were (72.25±24.81)%，(68.34±10.20)% and（8.87±5.18）×105 /mL,respectively.The cell viability and total number of cells in the three groups were not significantly different(P＞0.05).The cell purity of method Ⅱ was extremely significantly higher than that of methods Ⅰ and Ⅲ (Ρ＜0.01). The dynamic distribution of mast cells in the Percoll gradient separation liquid was that the cell viabilities of 3,4 layers were extremely significantly higher than 1 layer (Ρ＜0.01),significantly higher than 2 layer (Ρ＜0.05); the cell purity of 3 layer was extremely significantly higher than 1,4 layers (Ρ＜0.01), significantly higher than 2 layer (Ρ＜0.05); the number of cells of the 3,4 layers were extremely significantly higher than 1,2 layers (Ρ＜0.01). Therefore, adopting the method Ⅱ, layer 3 of extraction, the middle and lower 0.5 mL, which was close to the interface parts in 30%∶ 80% of the Percoll gradient separation liquid, the separation of mast cells of mice peritoneal cavity was best.

Key words: mice; mast cells; separation method

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