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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
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
Dit protocol biedt een methode om Mycobacterium tuberculosis infectie in menselijke M1- of M2-gepolariseerde macrofagen te bestuderen op basis van differentiatie van perifere bloed-monocyten naar macrofaag-achtige cellen die zijn geïnfecteerd met de GFP-gelabelde virulente stam H37Rv, en geanalyseerd met flowcytometrie met behulp van een 10-kleurenpaneel inclusief expressie van geselecteerde M1/M2 markers.
