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Polarisering af M1 og M2 Human Monocyte-afledte celler og analyse med Flow Cytometry på Mycobacterium tuberkulose Infektion
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Polarisering af M1 og M2 Human Monocyte-afledte celler og analyse med Flow Cytometry på Mycobacterium tuberkulose Infektion
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
Capítulos
- 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
Denne protokol giver en metode til at studere Mycobacterium tuberkulose infektion i humane M1- eller M2-polariserede makrofager baseret på differentiering af perifere blod-monocytter til makrofaglignende celler, der er inficeret med GFP-mærket virulent stamme H37Rv, og analyseret med flow cytometry ved hjælp af en 10-farve panel, herunder udtryk for udvalgte M1/M2 markører.
