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Quantification of Efferocytosis by Single-cell Fluorescence Microscopy
Published on: August 18, 2018
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In vitro Assessment of Efferocytic Capacity of Human Macrophages Using Flow Cytometry
Ana C G Salina1,2, Marlon Fortes-Rocha1, Larissa D Cunha1
1Departamento de Biologia Celular e Molecular e Bioagentes Patogênicos, Faculdade de Medicina de Ribeirão Preto, Universidade de São Paulo, Ribeirao Preto, Brazil.
Bio-Protocol
|December 29, 2023
Summary
This study presents a new flow cytometry assay to measure how efficiently human macrophages engulf apoptotic cells. This method models efferocytosis in vitro, aiding research into cell debris clearance.
Area of Science:
- Immunology
- Cell Biology
Background:
- Efferocytosis, the clearance of dying cells, is crucial for preventing cellular debris accumulation.
- Professional phagocytes perform efferocytosis, a process vital for tissue homeostasis.
- Existing methods for assessing efferocytosis include cell imaging and flow cytometry.
Purpose of the Study:
- To describe a novel in vitro assay for evaluating the uptake of apoptotic cells (ACs) by human macrophages.
- To provide a detailed protocol for preparing and labeling cells for efferocytosis assessment.
- To establish a model for studying the efficiency of AC engulfment in a continuous efferocytosis process.
Main Methods:
- Utilizing commercially available dyes to label human macrophages and apoptotic lymphocytes.
- Employing flow cytometry for quantitative analysis of apoptotic cell uptake.
- Developing an in vitro system to mimic continuous efferocytosis.
Main Results:
- Successfully established and detailed a protocol for assessing efferocytosis in vitro.
- Demonstrated the utility of flow cytometry for quantifying apoptotic cell engulfment by macrophages.
- Validated the assay based on previously published literature.
Conclusions:
- The described assay provides a reliable method for modeling efferocytosis in vitro.
- This protocol can be adapted to study the clearance of various cell types by different professional phagocytes.
- The assay facilitates research into the efficiency of apoptotic cell engulfment and its implications in biological processes.
