Please use this identifier to cite or link to this item: https://hdl.handle.net/2440/37461
Citations
Scopus Web of ScienceĀ® Altmetric
?
?
Type: Journal article
Title: Phagocytosis of apoptotic cells by human macrophages: analysis by multiparameter flow cytometry.
Author: Jersmann, H.
Ross, J.
Vivers, S.
Brown, S.
Haslett, C.
Dransfield, I.
Citation: Cytometry Part B: Clinical Cytometry, 2003; 51A(1):7-15
Publisher: Wiley-Liss
Issue Date: 2003
ISSN: 1552-4922
1552-4930
Statement of
Responsibility: 
Hubertus P. A. Jersmann, Katherine A. Ross, Sharon Vivers, Simon B. Brown, Christopher Haslett, Ian Dransfield
Abstract: <h4>Background</h4>Phagocytic removal of apoptotic cells is an important regulatory event in development, tissue homoeostasis, and inflammation. There are several methodologic problems with most in vitro studies of the molecular mechanisms of apoptotic cell phagocytosis. First, cell loss occurs during rigorous washing of adherent macrophages required to ensure removal of noningested particles. Second, discrimination of adherent or internalised apoptotic cells is difficult. Third, microscopic quantification is time consuming and has the potential for significant interobserver error. Fourth, subsequent analysis of phagocyte populations is difficult.<h4>Methods</h4>We used a flow cytometric method that allows quantification of phagocytosis of fluorescently labelled apoptotic cells with the use of multiparameter flow cytometric analysis.<h4>Results</h4>Phagocytosis of apoptotic cells was validated by use of inhibitors (cytochalasins) or low temperature and counterstaining with cell surface markers for the phagocytic targets to exclude binding to the phagocytic surface. Populations of phagocytic macrophages were sorted, and the presence of internalized apoptotic material was validated by microscopy.<h4>Conclusions</h4>The technique we used in this study allows observer-independent analysis of phagocytosis of apoptotic cells by macrophages. Importantly, phagocytic or nonphagocytic populations could be subjected to further characterization with the use of flow cytometry with additional fluorochrome reagents and can be re-cultured to study underlying regulatory mechanisms.
Keywords: Neutrophils
Macrophages
Humans
Cytochalasins
Fluoresceins
Antigens, Surface
Fluorescent Dyes
Flow Cytometry
Reproducibility of Results
Apoptosis
Phagocytosis
Description: The definitive version may be found at www.wiley.com
DOI: 10.1002/cyto.a.10005
Published version: http://www3.interscience.wiley.com/cgi-bin/abstract/102019594/ABSTRACT
Appears in Collections:Aurora harvest 6
Medicine publications

Files in This Item:
There are no files associated with this item.


Items in DSpace are protected by copyright, with all rights reserved, unless otherwise indicated.