Isolation of mouse alveolar macrophages by flow cytometry and identification

VernacularTitle:流式细胞术分选小鼠肺泡巨噬细胞及鉴定
Author: Xintao HAN ¹ ; Dianyuan ZHAO ; Li TANG
Author Information

1. 安徽医科大学
Keywords: alveolar macrophage; flow cytometry; real-time PCR; phagocytosis
From: Acta Universitatis Medicinalis Anhui 2017;52(11):1596-1600
CountryChina
Language:Chinese
Abstract: Objective To establish a method for isolating alveolar macrophage (AM) of mouse based on flow cy tometry.Methods The lungs were digested by collagen ⅣV in vitro to prepare single-cell suspension that was stained by CD11 b and CD1 1 c antibody.CD1 1 b1owCD1 1 c + cell population were AM and isolated by flow cytometry.After that,the cell viability was measured via the Typan blue staining,and the identification of AM was through flow cytometry and real-time PCR.Results CD1 1 b1owCD1 1 c + cell population was isolated by flow cytometry,the purity was (93 ± 2)％ and the cell viability was (80 ±5)％.The real-time PCR results showed that peroxisome proliferator-activated receptor γ (PPARγ) mRNA was highly expressed in AM isolated by flow cytometry (P ＜ 0.001).In addition,the functional assay showed that the isolated AM possess high phagocytic activity.Thus,the results described above demonstrate that the isolated cells were AM.Conclusion A method for obtaining AM based on flow cytometry was established.The method has high cell purity and good cell activity which can be used for functional experiments.