Journal article
A real time chemotaxis assay unveils unique migratory profiles amongst different primary murine macrophages
- 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.
- Publication status:
- Published
- Peer review status:
- Peer reviewed
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(Preview, Version of record, pdf, 1.7MB, Terms of use)
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- Publisher copy:
- 10.1371/journal.pone.0058744
Authors
- Publisher:
- Public Library of Science
- Journal:
- PLoS ONE More from this journal
- Volume:
- 8
- Issue:
- 3
- Pages:
- e58744
- Publication date:
- 2013-03-14
- Acceptance date:
- 2013-02-05
- DOI:
- EISSN:
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1932-6203
- ISSN:
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1932-6203
- Language:
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English
- Keywords:
- UUID:
-
uuid:c2248987-618c-426d-bd75-a8df5a32b9f6
- Local pid:
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pubs:390809
- Source identifiers:
-
390809
- Deposit date:
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2013-11-17
Terms of use
- Copyright holder:
- Iqbal et al
- Copyright date:
- 2013
- Notes:
- © 2013 Iqbal et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
- Licence:
- CC Attribution (CC BY)
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