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Polarization of M1 and M2 Human Monocyte-Derived Cells and Analysis with Flow Cytometry upon Mycobacterium tuberculosis Infection
JoVE Journal
Immunology and Infection
This content is Free Access.
JoVE Journal Immunology and Infection
Polarization of M1 and M2 Human Monocyte-Derived Cells and Analysis with Flow Cytometry upon Mycobacterium tuberculosis Infection
DOI:

10:43 min

September 18, 2020

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Chapters

  • 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

Automatic Translation

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.

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