Untouched isolation of functionally unaffected neutrophils from whole blood within 20 minutes
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1
Miltenyi Biotec GmbH, Germany
Human neutrophils are commonly obtained by density gradient centrifugation. Commonly used methods such as Ficoll-Paque gradients in combination with dextran sedimentation, discontinuous Percoll or Polymorphprep gradients are exceedingly time-consuming and frequently result in impure neutrophils which are contaminated by other leukocytes, mainly eosinophils.
With the new MACSxpress technology, untouched human leukocyte subsets can be isolated from up to 30 ml of anticoagulated whole blood. Erythrocyctes are aggregated and sedimented, while non-target cells are removed by immunomagnetic depletion with MACSxpress Beads, yielding untouched target cells of high purity.
MACSxpress-enriched neutrophils had average purities of 93% with yields of ~97% (n=14). ROS (reactive oxygen species) generation and expression levels of the surface markers CD11b, CD62L and CD181 (CXCR1) of MACSxpress-isolated neutrophils were comparable to cells isolated with density gradient centrifugation. In addition, separated neutrophils maintained efficient phagocytic capacity, which was assessed by uptake of opsonized bacteria and showed typical migration behavior toward fMLP. Cell viability was unaffected, as demonstrated by flow cytometric analysis of AnnexinV staining.
Keywords:
Neutrophils,
cell isolation,
innate immunity,
Granulocytes,
whole blood separation
Conference:
15th International Congress of Immunology (ICI), Milan, Italy, 22 Aug - 27 Aug, 2013.
Presentation Type:
Abstract
Topic:
Innate immunity
Citation:
Zyntek
C,
Schmitz
J and
Winkels
G
(2013). Untouched isolation of functionally unaffected neutrophils from whole blood within 20 minutes.
Front. Immunol.
Conference Abstract:
15th International Congress of Immunology (ICI).
doi: 10.3389/conf.fimmu.2013.02.00621
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Received:
12 Jun 2013;
Published Online:
22 Aug 2013.
*
Correspondence:
Mr. Gregor Winkels, Miltenyi Biotec GmbH, Bergisch Gladbach, Germany, Gregorw@miltenyibiotec.de