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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
Questo protocollo fornisce un metodo per studiare l'infezione da Mycobacterium tuberculosis nei macrofagi polarizzati M1 o M2 umani basati sulla differenziazione dei monociti del sangue periferico alle cellule simili a macrofagi che sono infettate dal ceppo virulento H37Rv etichettato GFP e analizzate con citometria a flusso utilizzando un pannello a 10 colori che include l'espressione di marcatori M1/M2 selezionati.
