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Immunology and Infection

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Polarization of M1 and M2 Human Monocyte-Derived Cells and Analysis with Flow Cytometry upon Mycobacterium tuberculosis Infection

Article DOI: 10.3791/61807-v • 10:43 min • September 18th, 2020
September 18th, 2020 •
1,2, 1, 1, 3, 1, 1,4, 1
1Center for Infectious Medicine (CIM), Department of Medicine Huddinge, ANA Futura, Karolinska Institutet, 2Infectious Diseases Division, International Centre for Diarrhoeal Disease Research, Bangladesh, 3Clinical Microbiology, Department of Laboratory Medicine (Labmed), ANA Futura, Karolinska Institutet, 4Childhood Cancer Research Unit, Department of Women's and Children's Health, Karolinska Institutet

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This protocol provides a method to study Mycobacterium tuberculosis infection in human M1- or M2-polarized macrophages based on differentiation of peripheral-blood-monocytes to macrophage-like cells that are infected with the GFP-labeled virulent strain H37Rv, and analyzed with flow cytometry using a 10-color panel including expression of selected M1/M2 markers.

Tags

Polarization M1 M2 Human Monocyte-derived Cells Flow Cytometry Mycobacterium Tuberculosis Infection Green Fluorescent Protein-labeled Mtb Macrophage Subsets 10 Color Flow Cytometry Phenotype Function PBMC Isolation Buffy Coats Density Gradient Medium Centrifugation Mononuclear Cell Layer Serum-free RPMI Medium Macrophage Differentiation Cell Culture Incubator
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