An Assay for Studying Fcγ Receptor-Driven Phagocytosis in Monocyte Monolayers
Published: January 31, 2024
Abstract
Source: Tong, T. N., et al. Use of a Monocyte Monolayer Assay to Evaluate Fcγ Receptor-mediated Phagocytosis. J. Vis. Exp. (2017)
This video showcases a monocyte monolayer assay utilizing primary monocytes from peripheral blood to assess Fcγ receptor-mediated phagocytosis with poly D antibody-opsonized R2R2 red blood cells.
Protocol
All procedures involving sample collection have been performed in accordance with the institute's IRB guidelines. 1. Peripheral Blood Mononuclear Cell Isolation Obtain human whole blood from a healthy donor or a patient via venipuncture using vacutainers containing acid-citrate-dextrose (ACD) anticoagulant (yellow-top tubes). NOTE: Whole blood can be stored in ACD at room temperature (18-22 °C) for up to 36 hours be…
Divulgaciones
The authors have nothing to disclose.
Materials
| RPMI 1640 | RPMI 1640 | R8758 | |
| Phosphate-buffered saline | Sigma | D8537 | |
| 8-chamber slides | Lab-Tek-ll | 154534 | |
| R2R2 (cDE/cDE) red blood cells | Commercially available (e.g., http:// www.bio-rad.com/en-ca/product/ reagent-red-blood-cells) | ||
| Polyclonal anti-D from human serum | Gamma Biologics | DIN 02247724 | Can be substituted with commercially available monoclonal anti-D or with Rh immune globulin |
| 100% methanol | Caledon | 6700-1-42 | |
| Cover slips | VWR | 48366 067 |
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An Assay for Studying Fcγ Receptor-Driven Phagocytosis in Monocyte Monolayers. J. Vis. Exp. (Pending Publication), e21912, doi: (2024).