The influence of different anticoagulants and sample preparation methods on measurement of mCD14 on bovine monocytes and polymorphonuclear neutrophil leukocytes.

BMC Research Notes
Eveline M Ibeagha-AwemuXin Zhao

Abstract

Membrane-CD14 (mCD14) is expressed on the surface of monocytes, macrophages and polymorphonuclear neutrophil leukocytes (PMN). mCD14 acts as a co-receptor along with Toll like receptor 4 (TLR 4) and MD-2 for the detection of lipopolysaccharide (LPS). However, studies using different sample preparation methods and anticoagulants have reported different levels of mCD14 on the surface of monocytes and neutrophils. In this study, the influence of various anticoagulants and processing methods on measurement of mCD14 on monocytes and neutrophils was examined. Whole blood samples were collected in vacutainer tubes containing either sodium heparin (HEPARIN), ethylenediaminetetraacetic acid (EDTA) or sodium citrate (CITRATE). mCD14 on neutrophils and monocytes in whole blood samples or isolated cells was measured by the method of flow cytometry using fluorescein isothiocyanate (FITC)-labeled monoclonal antibody. There was a significant difference (p < 0.05) in the mean channel fluorescence intensity (MFI) of mCD14 on neutrophils in whole blood samples anticoagulated with HEPARIN (MFI = 64.77) in comparison with those in whole blood samples anticoagulated with either EDTA (MFI = 38.25) or CITRATE (MFI = 43.7). The MFI of mCD14 on monocyt...Continue Reading

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Citations

Jun 21, 2012·Journal of Thrombosis and Haemostasis : JTH·M G BasavarajJ-B Hansen
Apr 15, 2014·Inflammation·Yuji TakedaHironobu Asao
Jul 28, 2018·Frontiers in Immunology·Shima Hassan BaienNicole de Buhr
Oct 19, 2020·Cytometry. Part a : the Journal of the International Society for Analytical Cytology·Paulina RybakowskaConcepción Marañón
Nov 11, 2020·Molecular Biology Reports·Jesse D ArmitageDino B A Tan
Jul 6, 2020·Methods : a Companion to Methods in Enzymology·F CecilianiC Lecchi

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Methods Mentioned

BETA
density gradient centrifugation
Flow Cytometry

Software Mentioned

Cell
Windows Multiple Document Interface for Flow Cytometry ( WinMDI )
Quest
SAS

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