Flow cytometric evaluation of a model for phagocytosis of cells undergoing apoptosis.
Phagocyte recognition of cells undergoing apoptosis is a rapid, efficient way of removing unwanted cells from tissue. The uptake of apoptotic cells prevents the release of potentially toxic cell contents that might otherwise damage neighbouring cells and elicit an inflammatory response. The aim of this work was to evaluate a simple cell culture assay to study phagocytosis of cells undergoing apoptosis. Fluorescent negatively charged beads (1 microm) or fluorescently labelled apoptotic cells, derived from etoposide-treated human monocytes (U937), were co-incubated with J774 cells or human peripheral blood macrophages for 1 h. Flow cytometry (FCM) showed an efficient uptake of both beads and apoptotic bodies. Phagocytosis of apoptotic cells but not of beads was significantly inhibited when macrophages were pre-incubated with cytochalasin D, suggesting that an experimental system based on beads is not an appropriate model of phagocytosis of apoptotic cells.
Schrijvers
Dorien M
DM
Division of Pharmacology, University of Antwerp, Universiteitsplein 1, Wilrijk B-2610, Belgium. dorien.shrijvers@ua.ac.be
Martinet
Wim
W
De Meyer
Guido R Y
GR
Andries
Luc
L
Herman
Arnold G
AG
Kockx
Mark M
MM
D000818:Animals; D017209:Apoptosis / Q000276:immunology*; D018929:Cell Culture Techniques / Q000379:methods; D015638:Cytochalasin D / Q000494:pharmacology; D005434:Flow Cytometry / Q000379:methods*; D005453:Fluorescence; D006801:Humans; D015151:Immunoblotting; D020287:In Situ Nick-End Labeling; D051379:Mice; D018613:Microscopy, Confocal; D008863:Microspheres; D009000:Monocytes / Q000187:drug effects / Q000276:immunology*; D019384:Nucleic Acid Synthesis Inhibitors / Q000494:pharmacology; D010587:Phagocytosis / Q000187:drug effects / Q000276:immunology*; D020298:U937 Cells
D019384:Nucleic Acid Synthesis Inhibitors; D015638:Cytochalasin D