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• W2150794310 abstract "Inappropriate neutrophil activation has been implicated in the pathology of several clinically important inflammatory conditions. Although murine models are extensively used in the investigation of such pathological processes, a reliable method by which viable, quiescent neutrophils can be isolated from murine blood has not been developed. Here we describe a novel method based on negative immunomagnetic separation, which yields highly pure populations of murine neutrophils. Blood is incubated with a cocktail of antibodies against specific cell markers on unwanted cells, and then with secondary antibody-coated magnetic beads. After running the preparation through a column within a magnetic field, labeled cells are retained, and a neutrophil-rich effluent is collected. This method yields a >95% pure suspension of >97% viable neutrophils, recovering approximately 70% of neutrophils from whole blood. Flow cytometric analysis shows little difference in surface L-selectin and CD18 expression on isolated neutrophils compared with neutrophils in whole blood, indicating that neutrophils are minimally activated bythe isolation process. Stimulation with phorbol 12-myristate 13-acetate (PMA) reduced L-selectin andincreased CD18 expression. Isolated neutrophilsmigrate under agarose in response to fMLP, and fluorescently labeled neutrophils transfused into recipient mice interact with postcapillary venules in a manner comparable to endogenous leukocytes. These findings show that neutrophils isolated using this method can be used for inflammatory studies in vitro and in vivo." @default.
• W2150794310 created "2016-06-24" @default.
• W2150794310 creator A5006903396 @default.
• W2150794310 creator A5029073857 @default.
• W2150794310 creator A5071605337 @default.
• W2150794310 creator A5078768064 @default.
• W2150794310 date "2001-08-01" @default.
• W2150794310 modified "2023-10-16" @default.
• W2150794310 title "A Novel Method for Isolation of Neutrophils from Murine Blood Using Negative Immunomagnetic Separation" @default.
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• W2150794310 doi "https://doi.org/10.1016/s0002-9440(10)61719-1" @default.
• W2150794310 hasPubMedCentralId "https://www.ncbi.nlm.nih.gov/pmc/articles/1850545" @default.
• W2150794310 hasPubMedId "https://pubmed.ncbi.nlm.nih.gov/11485906" @default.
• W2150794310 hasPublicationYear "2001" @default.
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