小鼠腹腔巨噬细胞吞噬乳胶颗粒实验条件筛选

Study on experimental conditions for phagocyteosis of mouse peritoneal macrophages

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 provide a reliable, sensitive and stable method for detecting the percentage of phagocytosis of mouse peritoneal macrophages. Methods The mouse peritoneal macrophages were incubated with latex beads in 24 well plate in cell culture incubator. The percentage of phagocytosis（PP） and phagocytosis index（PI） were detected by fluores- cence microscope. Results As the culture time increasing, PP of macrophages became higher （ P 〈 0. 05 ）. The PP of perito- neal macrophages increased with the increment of calf serum concentration. As the concentration of latex beads increasing, the PP of peritoneal macrophages also increased（ P 〈 0. 01 ）. The PP of peritoneal macrophages of adherent macrophages was higher than that of non-adherent macrophages（ P 〈 0. 01 ）. The PP of peritoneal macrophages of the Balb/c mice was signifi- cantly higher than that of the C57, KM and ICR mice （P 〈 0. 01 ）. Conclusion The optimal experimental conditions of the phagocyteosis of mouse peritoneal macrophages might be the concentration of latex beads of 1%o,the calf serum concentra- tion of 2.5% -5% ,adding the latex beads and continuous culturing for 2 hours at the time of 2 hours after the adherence of macrophages. Besides, it is indicated that Balb/c mouse is most suitable for detection of the capacity of phagocyteosis.