Development of a simple method for rapid isolation of polymorphonuclear leukocytes from human blood

Autor:	Robin C Minielly, Paul E. Bankey, Carol L. Miller-Graziano, Mita De, Asit De, Krzysztof Laudanski, Sarah E. Roach
Rok vydání:	2005
Předmět:	Time Factors Lipopolysaccharide medicine.drug_class Cell Survival Neutrophils Sialoglycoproteins Clinical Biochemistry Immunology Inflammation Cell Separation Biology Peripheral blood mononuclear cell Flow cytometry chemistry.chemical_compound Antigen Antigens CD medicine Immunology and Allergy Humans Secretion Cells Cultured Innate immune system medicine.diagnostic_test Receptor antagonist Flow Cytometry Antigens Differentiation Medical Laboratory Technology Interleukin 1 Receptor Antagonist Protein chemistry medicine.symptom Cell Adhesion Molecules
Zdroj:	Journal of immunoassayimmunochemistry. 26(1)
ISSN:	1532-1819
Popis:	Polymorphonuclear leukocytes (PMNs; commonly known as neutrophils) play essential roles in innate immunity and inflammation. Although there are standardized methods for the isolation of human neutrophils, they are time consuming and demand considerable technical expertise, making them unfeasible for many clinical applications. Here, we describe a simple and time-efficient technique for the isolation of human neutrophils, which adapts a readily available commercial cell preparation tube (CPT) currently in use for isolation of peripheral blood mononuclear cells (PBMC) and plasma and is now adapted to also yield neutrophils. The total time required for neutrophil isolation was less than 1 hr. Neutrophils isolated by this method were highly purified (> or =97%) as assessed by surface expression of the neutrophil specific marker, CD66b. Neutrophils isolated by this method were functional as demonstrated by their ability to secrete interleukin-1 receptor antagonist (IL-1RA). Neutrophils isolated using this new technique secreted significant amounts of soluble IL-1RA (929.3+/-197 pg/10(6)cells/mL) in response to lipopolysaccharide (LPS). Use of this adapted CPT method allows simultaneous isolation of functional human neutrophils as well as PBMC and plasma. Adoption of this new method will allow the conduct of different neutrophil assays at any clinical site without requiring trained laboratory personnel or a large staff time commitment.
Databáze:	OpenAIRE
Externí odkaz:	https://explore.openaire.eu/search/publication?articleId=doi_dedup___::bc508498fb3841a05cd2556da23007cc https://pubmed.ncbi.nlm.nih.gov/15754803 Zobrazit plný text záznamu Plný text ve formátu PDF Plný text ve formátu HTML
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