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目的建立和完善可靠、敏感、稳定的评价腹腔巨噬细胞吞噬功能方法。方法取小鼠腹腔巨噬细胞与乳胶颗粒在不同培养条件下混合后定量加入24孔板,37℃,5%CO2培养箱中孵育后,荧光显微镜下计数吞噬百分率和吞噬指数。结果随培养时间(0.5,1,2,4h)延长,腹腔巨噬细胞吞噬乳胶颗粒的吞噬百分率增加,不同培养时间的吞噬百分率之间差异均有统计学意义(P<0.05);随小牛血清浓度(2.5%,5%,10%)和乳胶颗粒浓度(0.25‰,0.5‰,1‰,2‰)增加,腹腔巨噬细胞吞噬百分率和吞噬指数升高,不同浓度小牛血清和乳胶颗粒下腹腔巨噬细胞吞噬百分率和吞噬指数之间差异均有统计学意义(P<0.01);贴壁处理后的腹腔巨噬细胞吞噬百分率和吞噬指数显著高于未经贴壁处理的细胞(P<0.01);在Balb/c、C57、昆明(KM)、ICR4种品系小鼠中,Balb/c小鼠腹腔巨噬细胞吞噬百分率显著高于其他品系小鼠(P<0.01)。结论乳胶颗粒浓度为1‰、血清浓度为2.5%～5%、细胞贴壁2h后吞噬培养2h,选用Balb/c小鼠为该实验的最佳实验条件。
Objective To establish and perfect a reliable, sensitive and stable method for evaluating phagocytic function of peritoneal macrophages. Methods Mouse peritoneal macrophages and latex particles were mixed under different culture conditions, and then added into 24-well plates. Incubated at 37 ℃ in 5% CO2 incubator, the phagocytic percentage and phagocytic index were counted under a fluorescence microscope. Results The phagocytosis rate of phagocytic latex particles in peritoneal macrophages increased with the prolongation of culture time (0.5, 1, 2, 4h), the difference of phagocytosis percentage in different culture time was statistically significant (P <0.05) Serum concentration (2.5%, 5%, 10%) and latex particle concentration (0.25 ‰, 0.5 ‰, 1 ‰, 2 ‰) increased peritoneal macrophages phagocytic percentage and phagocytic index increased with different concentrations of bovine serum and latex The difference of phagocytic percentage and phagocytic index between granulocyte peritoneal macrophages were statistically significant (P <0.01). The phagocytic percentage and phagocytic index of peritoneal macrophages after attachment treatment were significantly higher than those of non-adherent cells P <0.01). In Balb / c, C57, Kunming (KM) and ICR4 strains, the phagocytic percentage of peritoneal macrophages in Balb / c mice was significantly higher than that in other strains (P <0.01). Conclusion The latex particles concentration of 1 ‰, serum concentration of 2.5% to 5%, 2h after attachment of cells phagocytosis culture 2h, the selection of Balb / c mice for the experiment the best experimental conditions.