Document Detail


Highly purified selective isolation of eosinophils from human peripheral blood by negative immunomagnetic selection.
MedLine Citation:
PMID:  17406516     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Eosinophils are a minority constituent in human peripheral blood. The study of eosinophils has been limited by difficulty in achieving sufficient cell number and purity. We describe a modified protocol for immunomagnetic cell separation for efficient isolation of human peripheral blood eosinophils. We employ a mixture of monoclonal antibodies (mAbs) directed against cell-surface antigens on human hematopoietic cells combined with secondary labeling with a colloidal suspension of magnetic dextran-iron particles for negative selection of eosinophils. Unwanted labeled cells are retained in the magnetized column, permitting high recovery (70%) and purity (>98%) of eosinophils while retaining cell viability. Eosinophils remain quiescent after isolation, and stimulation caused by cytokines upregulates (i) cell-cell or cell-extracellular matrix protein adhesion, (ii) secretion of bioactive mediators and (iii) cell-surface adhesion molecules. This method for purified isolation is accomplished in < or = 4 h and preserves eosinophils in a quiescent, viable state.
Authors:
Nilda M Munoz; Alan R Leff
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Publication Detail:
Type:  Journal Article; Research Support, N.I.H., Extramural; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Nature protocols     Volume:  1     ISSN:  1750-2799     ISO Abbreviation:  Nat Protoc     Publication Date:  2006  
Date Detail:
Created Date:  2007-04-04     Completed Date:  2007-11-05     Revised Date:  2008-03-24    
Medline Journal Info:
Nlm Unique ID:  101284307     Medline TA:  Nat Protoc     Country:  England    
Other Details:
Languages:  eng     Pagination:  2613-20     Citation Subset:  IM    
Affiliation:
Section of Pulmonary and Critical Care Medicine, Department of Medicine and Pediatrics, The University of Chicago, Chicago, Illinois 60637, USA.
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MeSH Terms
Descriptor/Qualifier:
Eosinophils* / immunology
Humans
Immunomagnetic Separation / methods*
Grant Support
ID/Acronym/Agency:
HL-46368/HL/NHLBI NIH HHS; HL-56399/HL/NHLBI NIH HHS

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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