快速振荡法高效提取小鼠腹腔巨噬细胞研究  

Highly Effective Extraction of Mouse Peritoneal Macrophages by Quick Vibrating Method

作　　者：徐尤年[1] 刘桂林[1] 蒙臣 张诗海[1] 

机构地区：[1]华中科技大学同济医学院附属协和医院麻醉科,湖北省武汉市430032

出　　处：《中国全科医学》2016年第3期303-306,共4页Chinese General Practice

基　　金：国家自然科学基金青年基金资助项目(81201454)

摘　　要：目的通过与传统的腹腔巨噬细胞提取方法——轻揉法进行比较,建立一种高效提取小鼠腹腔巨噬细胞体外分离培养的方法——快速振荡法。方法 2014年11月选取雄性健康SPF级C57b1/6小鼠20只,采用随机数字表法分为轻揉组和振荡组,各10只。分别采用传统的腹部轻揉法和快速振荡法提取小鼠腹腔巨噬细胞。流式细胞仪检测巨噬细胞凋亡率,计算细胞总数及巨噬细胞总数。结果两组小鼠体质量和腹腔灌洗液回收率比较,差异无统计学意义（P〉0.05）。轻揉组巨噬细胞凋亡率为（5.44±0.08）%,振荡组巨噬细胞凋亡率为（5.60±0.12）%,差异无统计学意义（t=1.09,P=0.29）。振荡组细胞总数[（5.69±0.23）×10~6与（3.82±0.12）×10~6]和巨噬细胞总数[（2.64±0.08）×10~6与（1.66±0.07）×10~6]均多于轻揉组（P〈0.05）。结论快速振荡法提取小鼠腹腔巨噬细胞,巨噬细胞凋亡率无增加,细胞总数及巨噬细胞总数明显增加,是一种更加高效的提取腹腔巨噬细胞的方法。Objective To establish a high -efficiency method to obtain mouse peritoneal macrophages for separation and cultivation in vitro, which is quick vibrating method, by comparison with the traditional method, which is soft kneading method. Methods In November 2014, 20 male healthy C57bl/6 rats of SPF grade were collected, and random number table method was employed to divide the rats into soft kneading group and rapid oscillation group, with 10 rats in each group. Traditional abdomen soft kneading method and quick vibrating method were employed to extract mouse peritoneal macrophages. FCM was used to detect the apoptosis rate of peritoneal macrophages, and the total number of cells and peritoneal macrophages were calculated. Results The two groups were not significantly different in the body mass and the recovery rate of peritoneal lavage fluids （ P 〉0.05 ） . The apoptosis rate of peritoneal macrophages of soft kneading group was （5.44±0.08） % , and that of rapid oscillation group was （ 5.60 ±0. 12 ） %, without significant differences between the two groups. The rapid oscillation group was higher than the soft kneading group in the total number of cells [ （5.69 ± 0. 23 ） × 106 vs. （ 3.82 ± 0. 12） × 106 ] and peritoneal maerophages [ （ 2. 64±0. 08 ） × 106 vs. （ 1.66 ± 0. 07 ） ×106 ] （ P 〈 0.05 ） . Conclusion The extraction of peritoneal macrophages by the quick vibrating method cause no increase of the apoptosis rate of peritoneal macrophages and marked increase of the total number of cells and peritoneal macrophages. Therefore, it is a more effective method of peritoneal macrophages extraction.

关 键 词：巨噬细胞 小鼠 细胞分离 

分 类 号：R446.5[医药卫生—诊断学]