小鼠骨髓中性粒细胞分离纯化及活性检测研究

目的 探究小鼠骨髓中性粒细胞分离纯化，并进行鉴定以及活性的检测。方法 使用密度梯度离心法分离纯化小鼠骨髓中性粒细胞，得到的中性粒细胞采用免疫荧光法和高内涵分析仪检测中性粒细胞表面标志物Ly6G的表达。同时对得到的细胞加入佛波酯（phorbol 12-myristate 13-acetate，PMA）刺激后，采用免疫荧光的方法检测中性粒细胞释放髓过氧化物酶（myeloperoxidase，MPO）的表达情况。结果 用Histopaque^©-1077、Histopaque^©-1119分离纯化小鼠骨髓细胞后，细胞表达Ly6G，将分离纯化的细胞加入100 nmol/L PMA刺激2 h后检测MPO的表达增强，实验组是对照组的（3.26±0.22）倍。结论 该方法是一种简单、易操作的体外分离纯化、鉴定小鼠骨髓中性粒细胞的方法。

Separation,purification and identification of mouse bone marrow neutrophils

Objective For separation, purification and identification of mouse neutrophils from bone marrow. Methods Mouse bone marrow neutrophils were separated by density gradient centrifugation. Immunofluorescence and High Content Screening were used to detect the expression of Ly6G. Phorbol 12-myristate 13-acetate (PMA) was used to induce the expression of myeloperoxidase(MPO) released by neutrophils by immunofluorescent staining. Results After the mouse bone marrow cells were purified with Histopaque^©-1077, Histopaque^©-1119, the cells expressed Ly6G. High Content Screening was used to detect the expression of MPO, as demonstrated by the increased expression of MPO (3.26±0.22) stimulated by 100 nmol/L PMA for 2 hours. Conclusion It was a simple method for separation, purification and identification of mouse neutrophils from bone marrow.