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Polarization of M1 and M2 Human Monocyte-Derived Cells and Analysis with Flow Cytometry upon Mycobacterium tuberculosis Infection
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Immunologia e infezioni
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JoVE Journal Immunologia e infezioni
Polarization of M1 and M2 Human Monocyte-Derived Cells and Analysis with Flow Cytometry upon Mycobacterium tuberculosis Infection

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

DOI:
10.3791/61807-v

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10:43 min

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September 18, 2020

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, , , , , ,
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

Capitoli

  • 00:04Introduction
  • 00:44Peripheral Blood Mononuclear Cell (PBMC) Isolation
  • 01:40Monocyte-Derived Cell Differentiation and Polarization
  • 03:25Mycobacterium tuberculosis (Mtb) Culture
  • 05:00Monocyte-Derived Cell Mtb Infection
  • 06:03Flow Cytometry Staining and Analysis
  • 07:40Results: Representative Phenotypic M1 and M2 Cell Characterization Before and After Mtb Infection
  • 09:47Conclusion

Summary

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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

Keywords: M1 MacrophagesM2 MacrophagesMonocyte-derived CellsMycobacterium TuberculosisFlow CytometryPolarizationDifferentiationGM-CSFM-CSFIFN-gammaLPSIL-4

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