Please use this identifier to cite or link to this item: http://hdl.handle.net/2440/37461
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dc.contributor.authorJersmann, H.en
dc.contributor.authorRoss, J.en
dc.contributor.authorVivers, S.en
dc.contributor.authorBrown, S.en
dc.contributor.authorHaslett, C.en
dc.contributor.authorDransfield, I.en
dc.date.issued2003en
dc.identifier.citationCytometry Part A, 2003; 51A(1):7-15en
dc.identifier.issn1552-4922en
dc.identifier.issn1552-4930en
dc.identifier.urihttp://hdl.handle.net/2440/37461-
dc.descriptionThe definitive version may be found at www.wiley.comen
dc.description.abstractBACKGROUND: 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. METHODS: We used a flow cytometric method that allows quantification of phagocytosis of fluorescently labelled apoptotic cells with the use of multiparameter flow cytometric analysis. RESULTS: 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. CONCLUSIONS: 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.en
dc.description.statementofresponsibilityHubertus P. A. Jersmann, Katherine A. Ross, Sharon Vivers, Simon B. Brown, Christopher Haslett, Ian Dransfielden
dc.language.isoenen
dc.publisherWiley-Lissen
dc.source.urihttp://www3.interscience.wiley.com/cgi-bin/abstract/102019594/ABSTRACTen
dc.subjectNeutrophils; Macrophages; Humans; Cytochalasins; Fluoresceins; Antigens, Surface; Fluorescent Dyes; Flow Cytometry; Reproducibility of Results; Apoptosis; Phagocytosisen
dc.titlePhagocytosis of apoptotic cells by human macrophages: analysis by multiparameter flow cytometry.en
dc.typeJournal articleen
dc.identifier.rmid0020071026en
dc.identifier.doi10.1002/cyto.a.10005en
dc.identifier.pubid48676-
pubs.library.collectionMedicine publicationsen
pubs.verification-statusVerifieden
pubs.publication-statusPublisheden
dc.identifier.orcidJersmann, H. [0000-0003-1763-2736]en
Appears in Collections:Medicine publications

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