Document Detail


Isolation of bovine neutrophils with biomagnetic beads: comparison with standard Percoll density gradient isolation methods.
MedLine Citation:
PMID:  10410973     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
A prerequisite for studies on bovine neutrophils is a reliable method of neutrophil isolation from blood to obtain highly purified cell populations that are functionally active. Since current techniques of neutrophil isolation fall short of these requirements, we have developed a newer and more effective technique for isolation of bovine neutrophils that utilizes biomagnetic beads coated with a monoclonal antibody that recognizes an abundant surface antigen on bovine neutrophils to purify these cells. Comparison of the purity and viability of bovine neutrophils isolated by a conventional method (continuous Percoll density gradient) with this new method showed that neutrophils isolated with biomagnetic beads were higher in purity and had an increased yield. In addition, cells isolated with biomagnetic beads demonstrated normal or even improved function in assays of chemotaxis, phagocytosis, degranulation, and respiratory burst activity. Finally, bovine neutrophils isolated using this method showed an overall lower level of spontaneous apoptosis, which correlates well with the high level of viability observed in the purified cell preparations. Thus, this method represents a significant advance over current methods for isolating bovine neutrophils and would be widely applicable to labs studying the biochemistry and signal transduction pathways in these cells.
Authors:
J Soltys; S D Swain; K M Sipes; L K Nelson; A J Hanson; J M Kantele; M A Jutila; M T Quinn
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Publication Detail:
Type:  Comparative Study; Journal Article; Research Support, Non-U.S. Gov't; Research Support, U.S. Gov't, Non-P.H.S.; Research Support, U.S. Gov't, P.H.S.    
Journal Detail:
Title:  Journal of immunological methods     Volume:  226     ISSN:  0022-1759     ISO Abbreviation:  J. Immunol. Methods     Publication Date:  1999 Jun 
Date Detail:
Created Date:  1999-08-13     Completed Date:  1999-08-13     Revised Date:  2007-11-14    
Medline Journal Info:
Nlm Unique ID:  1305440     Medline TA:  J Immunol Methods     Country:  NETHERLANDS    
Other Details:
Languages:  eng     Pagination:  71-84     Citation Subset:  IM    
Affiliation:
Department of Veterinary Molecular Biology, Montana State University, Bozeman 59717, USA.
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MeSH Terms
Descriptor/Qualifier:
Animals
Antibodies, Monoclonal / immunology
Antibody Specificity
Antigens, CD18 / biosynthesis
Apoptosis
Cattle
Cell Degranulation
Centrifugation, Density Gradient / methods*
Chemotaxis
Immunomagnetic Separation / methods*
L-Selectin / biosynthesis
Mice
Mice, Inbred BALB C
Neutrophils* / cytology,  immunology,  metabolism,  physiology
Phagocytosis
Povidone
Respiratory Burst
Silicon Dioxide
Grant Support
ID/Acronym/Agency:
S10 RR11877/RR/NCRR NIH HHS
Chemical
Reg. No./Substance:
0/Antibodies, Monoclonal; 0/Antigens, CD18; 126880-86-2/L-Selectin; 65455-52-9/Percoll; 7631-86-9/Silicon Dioxide; 9003-39-8/Povidone

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


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