A Real Time Chemotaxis Assay Unveils Unique Migratory Profiles amongst Different Primary Murine Macrophages
Open Access
- 14 March 2013
- journal article
- research article
- Published by Public Library of Science (PLoS) in PLOS ONE
- Vol. 8 (3), e58744
- https://doi.org/10.1371/journal.pone.0058744
Abstract
Chemotaxis assays are an invaluable tool for studying the biological activity of inflammatory mediators such as CC chemokines, which have been implicated in a wide range of chronic inflammatory diseases. Conventional chemotaxis systems such as the modified Boyden chamber are limited in terms of the data captured given that the assays are analysed at a single time-point. We report the optimisation and validation of a label-free, real-time cell migration assay based on electrical cell impedance to measure chemotaxis of different primary murine macrophage populations in response to a range of CC chemokines and other chemoattractant signalling molecules. We clearly demonstrate key differences in the migratory behavior of different murine macrophage populations and show that this dynamic system measures true macrophage chemotaxis rather than chemokinesis or fugetaxis. We highlight an absolute requirement for Gαi signaling and actin cytoskeletal rearrangement as demonstrated by Pertussis toxin and cytochalasin D inhibition. We also studied the chemotaxis of CD14+ human monocytes and demonstrate distinct chemotactic profiles amongst different monocyte donors to CCL2. This real-time chemotaxis assay will allow a detailed analysis of factors that regulate macrophage responses to chemoattractant cytokines and inflammatory mediators.Keywords
This publication has 36 references indexed in Scilit:
- The neuroimmune guidance cue netrin-1 promotes atherosclerosis by inhibiting the emigration of macrophages from plaquesNature Immunology, 2012
- Site-Directed Mutagenesis of the CC Chemokine Binding Protein 35K-Fc Reveals Residues Essential for Activity and Mutations That Increase the Potency of CC Chemokine BlockadeMolecular Pharmacology, 2011
- Effect of CC Chemokine Receptor 2 CCR2 Blockade on Serum C-Reactive Protein in Individuals at Atherosclerotic Risk and With a Single Nucleotide Polymorphism of the Monocyte Chemoattractant Protein-1 Promoter RegionThe American Journal of Cardiology, 2011
- A Method for Generation of Bone Marrow-Derived Macrophages from Cryopreserved Mouse Bone Marrow CellsPLOS ONE, 2010
- Synthetic chemerin-derived peptides suppress inflammation through ChemR23The Journal of Experimental Medicine, 2008
- The macrophage: Past, present and futureEuropean Journal of Immunology, 2007
- Macrophage heterogeneity and tissue lipidsJCI Insight, 2007
- Chronic inflammation: a failure of resolution?International Journal of Experimental Pathology, 2006
- Resolution of inflammation: the beginning programs the endNature Immunology, 2005
- Resolution of inflammationInternational Journal of Immunopharmacology, 2000