三种方法提取大鼠原代巨噬细胞的生物学特点比较

Comparison of the biological characteristics of primary rat macrophages extracted by three methods

背景:提取原代大鼠巨噬细胞的方法包括肺泡灌洗液提取、脾脏提取及外周血提取等多种方法,这些方法各有优缺点,巨噬细胞活性、状态及数量各有不同。目的:分析3种不同组织来源的巨噬细胞原代提取及培养的较优选择。方法:在同一实验条件下,采用肺泡灌洗液提取、脾脏提取及外周血提取3种不同的原代细胞培养方法培养大鼠巨噬细胞。结果与结论:(1)肺泡灌洗液提取、脾脏提取及外周血提取法各组巨噬细胞培养提取细胞个数平均值分别为(1.14±0.14)×10~6个、(9.29±0.19)×10~6个及(3.17±0.13)×10~6个,3组间比较差异有显著性意义(P<0.001);(2)3种方法提取细胞存活的平均时间分别为(111.00±5.98),(100.00±4.60)及(97.00±3.43)h,差异有显著性意义(P<0.001);(3)3种方法提取细胞的活性比较,肺泡灌洗液提取出的巨噬细胞显著高于另2组(P<0.05),而脾脏提取与外周血提取差异无显著性意义(P>0.05);(4)结果提示,3种方法中,肺泡灌洗液提取的巨噬细胞活性最强,脾脏提取的巨噬细胞数量最多。

BACKGROUND： Methods for extracting primary macrophages from rats include bronchoalveolar lavage fluid（BALF） extraction, spleen extraction, and peripheral blood extraction. These methods have their own advantages and disadvantages, and the cells show different activities, status and quantities. OBJECTIVE： To analyze the better selection of primary extraction and culture of macrophages from three different tissue sources. METHODS： Under the same experimental conditions, primary rat macrophages were cultured using three different methods： BALF extraction, spleen extraction and peripheral blood extraction. RESULTS AND CONCLUSION： The average numbers of primary macrophage extracted from BALF, spleen and peripheral blood were（1.14±0.14）×10^6,（9.29±0.19）×10^6 and（3.17±0.13）×10^6 respectively, there was a significant difference among groups（P〈0.001）. The average survival time of the three methods for cell extraction was（111.00±5.98）,（100.00±4.60） and（97.00±3.43） hours respectively, the difference was significant（P〈0.001）. The activity of cells extracted from BALF was significantly higher than that of the other two tissues（P〈0.05）, but there was no significant difference between spleen and peripheral blood（P〈0.05）. These results indicate that BALF extracts the most activated macrophage can be extracted in BALF, and the largest number of macrophages can be extracted from the spleen.