Analysis of Myeloid Cells in Mouse Tissues with Flow Cytometry

Abstract

Summary Myeloid cells, including dendritic cells (DCs), granulocytes, monocytes, monocyte-derived cells and macrophages, are important players in the immune response, but their identification is not as clear as lymphocytes, especially in tissues. This protocol details the step-by-step procedure for the analysis of myeloid populations in various mouse tissues by flow cytometry. For complete details on the use and execution of this protocol, please refer to Liu et al. (2019) .

A novel multikinase inhibitor SKLB‐YTH‐60 ameliorates inflammation and fibrosis in bleomycin‐induced lung fibrosis mouse models

“…We analysed the percentage of MDSCs and macrophages by Flow cytometry (FCM). 22 Use scissors to cut approximately 30 mg of mouse lung tissues into small pieces and use 5 mL of collagenase-IV with a concentration of 1 mg•mL −1 , digested at 37°C for at least 1 hour. Centrifuge at 400 g for 5 minutes and discard the supernatant.…”

The SARS-CoV-2 receptor-binding domain facilitates neutrophil transepithelial migration and nanoparticle uptake in the mice airways

“…To investigate the influence of systemic ACE2 inhibition on the neutrophil-mediated response, we injected mice with selective ACE2 inhibitor MLN-4760, which was shown to affect mouse ACE2 (Ye et al, 2012). Using the strategy suggested by Liu et al (Liu et al, 2020) with slight modifications, we estimated the myeloid cell and neutrophil proportions in the periphery blood of mice that received MLN-4760 and control mice (Fig. 2A,B).…”

“…For the flow cytometry analysis, we used the strategy for the peripheral blood myeloid cell detection, which was recommended by Liu et al (Liu et al, 2020). The samples were preincubated with anti-mouse CD16/CD32 (Miltenyi Biotec, 130-092-574) for 15 min.…”

Decursinol Angelate Mitigates Sepsis Induced by Methicillin-Resistant Staphylococcus aureus Infection by Modulating the Inflammatory Responses of Macrophages

“…A single cell suspension was prepared as described elsewhere [38]. The liver, lung and kidneys were harvested after cardiac perfusion and cut into pieces followed by incubation at 37 • C for 30 min with the following digestion buffers: 25 µg/mL of Liberase DL, 25 µg/mL of Liberase TL (Roche, Basel, Switzerland) and 125 µg/mL of DNase I (Sigma, St. Louis, MO, USA) in serum-free RPMI media.…”