Un abonnement à JoVE est nécessaire pour voir ce contenu. Connectez-vous ou commencez votre essai gratuit.
This video demonstrates in vitro stimulation of macrophage extracellular traps (METs) in human monocyte-derive macrophages using different inflammatory stimuli such as TNF-alpha, PMA, and HOCL. The video also demonstrates the method of SYTOX dye staining of METs for visualization.
1. Polarization of HMDM
Access restricted. Please log in or start a trial to view this content.
Name | Company | Catalog Number | Comments |
120Q broad spectrum fluorescent light source | EXFO Photonic Solutions, Toronto, Canada | x-cite series | |
Corning CellBIND Multiple Well Plate (12 wells) | Sigma-Aldrich | CLS3336 | For cell culture |
Hypochlorous acid (HOCl) | Sigma-Aldrich | 320331 | For MET stimulation |
Interferon gamma | Thermo-Fisher | PMC4031 | For M1 priming |
Interleukin 4 | Integrated Sciences | rhil-4 | For M2 priming |
Interleukin 8 | Miltenyl Biotec | 130-093-943 | For MET stimulation |
L-Glutamine | Sigma-Aldrich | 59202C | Added to culture media |
Olympus IX71 inverted microscope | Olympus, Tokyo, Japan | ||
Phorbol 12- myristate 13-acetate (PMA) | Sigma-Aldrich | P8139 | For MET stimulation |
Phosphate buffered saline (PBS) | Sigma-Aldrich | D5652 | For washing steps |
RPMI-1640 media | Sigma-Aldrich | R8758 | For cell culture |
SYTOX green | Life Technologies | S7020 | For MET visulaization |
TH4-200 brightfield light source | Olympus, Tokyo, Japan | x-cite series | |
Tumor necrosis factor alpha | Lonza | 300-01A-50 | For MET stimulation |
Access restricted. Please log in or start a trial to view this content.
This article has been published
Video Coming Soon
Source: Zhang, Y., et al. In Vitro Stimulation and Visualization of Extracellular Trap Release in Differentiated Human Monocyte-derived Macrophages. J. Vis. Exp. (2019).