A real time chemotaxis assay unveils unique migratory profiles amongst different primary murine macrophages.

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...
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Autor*in:	Asif J Iqbal [verfasserIn] Daniel Regan-Komito [verfasserIn] Ivy Christou [verfasserIn] Gemma E White [verfasserIn] Eileen McNeill [verfasserIn] Amy Kenyon [verfasserIn] Lewis Taylor [verfasserIn] Theodore S Kapellos [verfasserIn] Edward A Fisher [verfasserIn] Keith M Channon [verfasserIn] David R Greaves [verfasserIn]

Format:	E-Artikel
Sprache:	Englisch

Erschienen:	2013

Übergeordnetes Werk:	In: PLoS ONE - Public Library of Science (PLoS), 2007, 8(2013), 3, p e58744
Übergeordnetes Werk:	volume:8 ; year:2013 ; number:3, p e58744

Links:	Link aufrufen Link aufrufen Link aufrufen Journal toc

DOI / URN:	10.1371/journal.pone.0058744

Katalog-ID:	DOAJ054627397

Internformat


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publishDate	2013
allfields	10.1371/journal.pone.0058744 doi (DE-627)DOAJ054627397 (DE-599)DOAJfce6674479494768832c2d37a8acafe2 DE-627 ger DE-627 rakwb eng Asif J Iqbal verfasserin aut A real time chemotaxis assay unveils unique migratory profiles amongst different primary murine macrophages. 2013 Text txt rdacontent Computermedien c rdamedia Online-Ressource cr rdacarrier 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. Medicine R Science Q Daniel Regan-Komito verfasserin aut Ivy Christou verfasserin aut Gemma E White verfasserin aut Eileen McNeill verfasserin aut Amy Kenyon verfasserin aut Lewis Taylor verfasserin aut Theodore S Kapellos verfasserin aut Edward A Fisher verfasserin aut Keith M Channon verfasserin aut David R Greaves verfasserin aut In PLoS ONE Public Library of Science (PLoS), 2007 8(2013), 3, p e58744 (DE-627)523574592 (DE-600)2267670-3 19326203 nnns volume:8 year:2013 number:3, p e58744 https://doi.org/10.1371/journal.pone.0058744 kostenfrei https://doaj.org/article/fce6674479494768832c2d37a8acafe2 kostenfrei http://europepmc.org/articles/PMC3597586?pdf=render kostenfrei https://doaj.org/toc/1932-6203 Journal toc kostenfrei GBV_USEFLAG_A SYSFLAG_A GBV_DOAJ GBV_ILN_11 GBV_ILN_20 GBV_ILN_22 GBV_ILN_23 GBV_ILN_24 GBV_ILN_31 GBV_ILN_34 GBV_ILN_39 GBV_ILN_40 GBV_ILN_60 GBV_ILN_62 GBV_ILN_63 GBV_ILN_65 GBV_ILN_69 GBV_ILN_70 GBV_ILN_73 GBV_ILN_74 GBV_ILN_95 GBV_ILN_105 GBV_ILN_110 GBV_ILN_151 GBV_ILN_161 GBV_ILN_170 GBV_ILN_171 GBV_ILN_206 GBV_ILN_213 GBV_ILN_224 GBV_ILN_230 GBV_ILN_235 GBV_ILN_285 GBV_ILN_293 GBV_ILN_370 GBV_ILN_602 GBV_ILN_702 GBV_ILN_2001 GBV_ILN_2003 GBV_ILN_2005 GBV_ILN_2006 GBV_ILN_2008 GBV_ILN_2009 GBV_ILN_2010 GBV_ILN_2011 GBV_ILN_2014 GBV_ILN_2015 GBV_ILN_2020 GBV_ILN_2021 GBV_ILN_2025 GBV_ILN_2031 GBV_ILN_2038 GBV_ILN_2044 GBV_ILN_2048 GBV_ILN_2050 GBV_ILN_2055 GBV_ILN_2056 GBV_ILN_2057 GBV_ILN_2061 GBV_ILN_2111 GBV_ILN_2113 GBV_ILN_2190 GBV_ILN_2522 GBV_ILN_4012 GBV_ILN_4037 GBV_ILN_4112 GBV_ILN_4125 GBV_ILN_4126 GBV_ILN_4249 GBV_ILN_4305 GBV_ILN_4306 GBV_ILN_4307 GBV_ILN_4313 GBV_ILN_4322 GBV_ILN_4323 GBV_ILN_4324 GBV_ILN_4325 GBV_ILN_4335 GBV_ILN_4338 GBV_ILN_4367 GBV_ILN_4700 AR 8 2013 3, p e58744
spelling	10.1371/journal.pone.0058744 doi (DE-627)DOAJ054627397 (DE-599)DOAJfce6674479494768832c2d37a8acafe2 DE-627 ger DE-627 rakwb eng Asif J Iqbal verfasserin aut A real time chemotaxis assay unveils unique migratory profiles amongst different primary murine macrophages. 2013 Text txt rdacontent Computermedien c rdamedia Online-Ressource cr rdacarrier 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. Medicine R Science Q Daniel Regan-Komito verfasserin aut Ivy Christou verfasserin aut Gemma E White verfasserin aut Eileen McNeill verfasserin aut Amy Kenyon verfasserin aut Lewis Taylor verfasserin aut Theodore S Kapellos verfasserin aut Edward A Fisher verfasserin aut Keith M Channon verfasserin aut David R Greaves verfasserin aut In PLoS ONE Public Library of Science (PLoS), 2007 8(2013), 3, p e58744 (DE-627)523574592 (DE-600)2267670-3 19326203 nnns volume:8 year:2013 number:3, p e58744 https://doi.org/10.1371/journal.pone.0058744 kostenfrei https://doaj.org/article/fce6674479494768832c2d37a8acafe2 kostenfrei http://europepmc.org/articles/PMC3597586?pdf=render kostenfrei https://doaj.org/toc/1932-6203 Journal toc kostenfrei GBV_USEFLAG_A SYSFLAG_A GBV_DOAJ GBV_ILN_11 GBV_ILN_20 GBV_ILN_22 GBV_ILN_23 GBV_ILN_24 GBV_ILN_31 GBV_ILN_34 GBV_ILN_39 GBV_ILN_40 GBV_ILN_60 GBV_ILN_62 GBV_ILN_63 GBV_ILN_65 GBV_ILN_69 GBV_ILN_70 GBV_ILN_73 GBV_ILN_74 GBV_ILN_95 GBV_ILN_105 GBV_ILN_110 GBV_ILN_151 GBV_ILN_161 GBV_ILN_170 GBV_ILN_171 GBV_ILN_206 GBV_ILN_213 GBV_ILN_224 GBV_ILN_230 GBV_ILN_235 GBV_ILN_285 GBV_ILN_293 GBV_ILN_370 GBV_ILN_602 GBV_ILN_702 GBV_ILN_2001 GBV_ILN_2003 GBV_ILN_2005 GBV_ILN_2006 GBV_ILN_2008 GBV_ILN_2009 GBV_ILN_2010 GBV_ILN_2011 GBV_ILN_2014 GBV_ILN_2015 GBV_ILN_2020 GBV_ILN_2021 GBV_ILN_2025 GBV_ILN_2031 GBV_ILN_2038 GBV_ILN_2044 GBV_ILN_2048 GBV_ILN_2050 GBV_ILN_2055 GBV_ILN_2056 GBV_ILN_2057 GBV_ILN_2061 GBV_ILN_2111 GBV_ILN_2113 GBV_ILN_2190 GBV_ILN_2522 GBV_ILN_4012 GBV_ILN_4037 GBV_ILN_4112 GBV_ILN_4125 GBV_ILN_4126 GBV_ILN_4249 GBV_ILN_4305 GBV_ILN_4306 GBV_ILN_4307 GBV_ILN_4313 GBV_ILN_4322 GBV_ILN_4323 GBV_ILN_4324 GBV_ILN_4325 GBV_ILN_4335 GBV_ILN_4338 GBV_ILN_4367 GBV_ILN_4700 AR 8 2013 3, p e58744
allfields_unstemmed	10.1371/journal.pone.0058744 doi (DE-627)DOAJ054627397 (DE-599)DOAJfce6674479494768832c2d37a8acafe2 DE-627 ger DE-627 rakwb eng Asif J Iqbal verfasserin aut A real time chemotaxis assay unveils unique migratory profiles amongst different primary murine macrophages. 2013 Text txt rdacontent Computermedien c rdamedia Online-Ressource cr rdacarrier 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. Medicine R Science Q Daniel Regan-Komito verfasserin aut Ivy Christou verfasserin aut Gemma E White verfasserin aut Eileen McNeill verfasserin aut Amy Kenyon verfasserin aut Lewis Taylor verfasserin aut Theodore S Kapellos verfasserin aut Edward A Fisher verfasserin aut Keith M Channon verfasserin aut David R Greaves verfasserin aut In PLoS ONE Public Library of Science (PLoS), 2007 8(2013), 3, p e58744 (DE-627)523574592 (DE-600)2267670-3 19326203 nnns volume:8 year:2013 number:3, p e58744 https://doi.org/10.1371/journal.pone.0058744 kostenfrei https://doaj.org/article/fce6674479494768832c2d37a8acafe2 kostenfrei http://europepmc.org/articles/PMC3597586?pdf=render kostenfrei https://doaj.org/toc/1932-6203 Journal toc kostenfrei GBV_USEFLAG_A SYSFLAG_A GBV_DOAJ GBV_ILN_11 GBV_ILN_20 GBV_ILN_22 GBV_ILN_23 GBV_ILN_24 GBV_ILN_31 GBV_ILN_34 GBV_ILN_39 GBV_ILN_40 GBV_ILN_60 GBV_ILN_62 GBV_ILN_63 GBV_ILN_65 GBV_ILN_69 GBV_ILN_70 GBV_ILN_73 GBV_ILN_74 GBV_ILN_95 GBV_ILN_105 GBV_ILN_110 GBV_ILN_151 GBV_ILN_161 GBV_ILN_170 GBV_ILN_171 GBV_ILN_206 GBV_ILN_213 GBV_ILN_224 GBV_ILN_230 GBV_ILN_235 GBV_ILN_285 GBV_ILN_293 GBV_ILN_370 GBV_ILN_602 GBV_ILN_702 GBV_ILN_2001 GBV_ILN_2003 GBV_ILN_2005 GBV_ILN_2006 GBV_ILN_2008 GBV_ILN_2009 GBV_ILN_2010 GBV_ILN_2011 GBV_ILN_2014 GBV_ILN_2015 GBV_ILN_2020 GBV_ILN_2021 GBV_ILN_2025 GBV_ILN_2031 GBV_ILN_2038 GBV_ILN_2044 GBV_ILN_2048 GBV_ILN_2050 GBV_ILN_2055 GBV_ILN_2056 GBV_ILN_2057 GBV_ILN_2061 GBV_ILN_2111 GBV_ILN_2113 GBV_ILN_2190 GBV_ILN_2522 GBV_ILN_4012 GBV_ILN_4037 GBV_ILN_4112 GBV_ILN_4125 GBV_ILN_4126 GBV_ILN_4249 GBV_ILN_4305 GBV_ILN_4306 GBV_ILN_4307 GBV_ILN_4313 GBV_ILN_4322 GBV_ILN_4323 GBV_ILN_4324 GBV_ILN_4325 GBV_ILN_4335 GBV_ILN_4338 GBV_ILN_4367 GBV_ILN_4700 AR 8 2013 3, p e58744
allfieldsGer	10.1371/journal.pone.0058744 doi (DE-627)DOAJ054627397 (DE-599)DOAJfce6674479494768832c2d37a8acafe2 DE-627 ger DE-627 rakwb eng Asif J Iqbal verfasserin aut A real time chemotaxis assay unveils unique migratory profiles amongst different primary murine macrophages. 2013 Text txt rdacontent Computermedien c rdamedia Online-Ressource cr rdacarrier 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. Medicine R Science Q Daniel Regan-Komito verfasserin aut Ivy Christou verfasserin aut Gemma E White verfasserin aut Eileen McNeill verfasserin aut Amy Kenyon verfasserin aut Lewis Taylor verfasserin aut Theodore S Kapellos verfasserin aut Edward A Fisher verfasserin aut Keith M Channon verfasserin aut David R Greaves verfasserin aut In PLoS ONE Public Library of Science (PLoS), 2007 8(2013), 3, p e58744 (DE-627)523574592 (DE-600)2267670-3 19326203 nnns volume:8 year:2013 number:3, p e58744 https://doi.org/10.1371/journal.pone.0058744 kostenfrei https://doaj.org/article/fce6674479494768832c2d37a8acafe2 kostenfrei http://europepmc.org/articles/PMC3597586?pdf=render kostenfrei https://doaj.org/toc/1932-6203 Journal toc kostenfrei GBV_USEFLAG_A SYSFLAG_A GBV_DOAJ GBV_ILN_11 GBV_ILN_20 GBV_ILN_22 GBV_ILN_23 GBV_ILN_24 GBV_ILN_31 GBV_ILN_34 GBV_ILN_39 GBV_ILN_40 GBV_ILN_60 GBV_ILN_62 GBV_ILN_63 GBV_ILN_65 GBV_ILN_69 GBV_ILN_70 GBV_ILN_73 GBV_ILN_74 GBV_ILN_95 GBV_ILN_105 GBV_ILN_110 GBV_ILN_151 GBV_ILN_161 GBV_ILN_170 GBV_ILN_171 GBV_ILN_206 GBV_ILN_213 GBV_ILN_224 GBV_ILN_230 GBV_ILN_235 GBV_ILN_285 GBV_ILN_293 GBV_ILN_370 GBV_ILN_602 GBV_ILN_702 GBV_ILN_2001 GBV_ILN_2003 GBV_ILN_2005 GBV_ILN_2006 GBV_ILN_2008 GBV_ILN_2009 GBV_ILN_2010 GBV_ILN_2011 GBV_ILN_2014 GBV_ILN_2015 GBV_ILN_2020 GBV_ILN_2021 GBV_ILN_2025 GBV_ILN_2031 GBV_ILN_2038 GBV_ILN_2044 GBV_ILN_2048 GBV_ILN_2050 GBV_ILN_2055 GBV_ILN_2056 GBV_ILN_2057 GBV_ILN_2061 GBV_ILN_2111 GBV_ILN_2113 GBV_ILN_2190 GBV_ILN_2522 GBV_ILN_4012 GBV_ILN_4037 GBV_ILN_4112 GBV_ILN_4125 GBV_ILN_4126 GBV_ILN_4249 GBV_ILN_4305 GBV_ILN_4306 GBV_ILN_4307 GBV_ILN_4313 GBV_ILN_4322 GBV_ILN_4323 GBV_ILN_4324 GBV_ILN_4325 GBV_ILN_4335 GBV_ILN_4338 GBV_ILN_4367 GBV_ILN_4700 AR 8 2013 3, p e58744
allfieldsSound	10.1371/journal.pone.0058744 doi (DE-627)DOAJ054627397 (DE-599)DOAJfce6674479494768832c2d37a8acafe2 DE-627 ger DE-627 rakwb eng Asif J Iqbal verfasserin aut A real time chemotaxis assay unveils unique migratory profiles amongst different primary murine macrophages. 2013 Text txt rdacontent Computermedien c rdamedia Online-Ressource cr rdacarrier 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. Medicine R Science Q Daniel Regan-Komito verfasserin aut Ivy Christou verfasserin aut Gemma E White verfasserin aut Eileen McNeill verfasserin aut Amy Kenyon verfasserin aut Lewis Taylor verfasserin aut Theodore S Kapellos verfasserin aut Edward A Fisher verfasserin aut Keith M Channon verfasserin aut David R Greaves verfasserin aut In PLoS ONE Public Library of Science (PLoS), 2007 8(2013), 3, p e58744 (DE-627)523574592 (DE-600)2267670-3 19326203 nnns volume:8 year:2013 number:3, p e58744 https://doi.org/10.1371/journal.pone.0058744 kostenfrei https://doaj.org/article/fce6674479494768832c2d37a8acafe2 kostenfrei http://europepmc.org/articles/PMC3597586?pdf=render kostenfrei https://doaj.org/toc/1932-6203 Journal toc kostenfrei GBV_USEFLAG_A SYSFLAG_A GBV_DOAJ GBV_ILN_11 GBV_ILN_20 GBV_ILN_22 GBV_ILN_23 GBV_ILN_24 GBV_ILN_31 GBV_ILN_34 GBV_ILN_39 GBV_ILN_40 GBV_ILN_60 GBV_ILN_62 GBV_ILN_63 GBV_ILN_65 GBV_ILN_69 GBV_ILN_70 GBV_ILN_73 GBV_ILN_74 GBV_ILN_95 GBV_ILN_105 GBV_ILN_110 GBV_ILN_151 GBV_ILN_161 GBV_ILN_170 GBV_ILN_171 GBV_ILN_206 GBV_ILN_213 GBV_ILN_224 GBV_ILN_230 GBV_ILN_235 GBV_ILN_285 GBV_ILN_293 GBV_ILN_370 GBV_ILN_602 GBV_ILN_702 GBV_ILN_2001 GBV_ILN_2003 GBV_ILN_2005 GBV_ILN_2006 GBV_ILN_2008 GBV_ILN_2009 GBV_ILN_2010 GBV_ILN_2011 GBV_ILN_2014 GBV_ILN_2015 GBV_ILN_2020 GBV_ILN_2021 GBV_ILN_2025 GBV_ILN_2031 GBV_ILN_2038 GBV_ILN_2044 GBV_ILN_2048 GBV_ILN_2050 GBV_ILN_2055 GBV_ILN_2056 GBV_ILN_2057 GBV_ILN_2061 GBV_ILN_2111 GBV_ILN_2113 GBV_ILN_2190 GBV_ILN_2522 GBV_ILN_4012 GBV_ILN_4037 GBV_ILN_4112 GBV_ILN_4125 GBV_ILN_4126 GBV_ILN_4249 GBV_ILN_4305 GBV_ILN_4306 GBV_ILN_4307 GBV_ILN_4313 GBV_ILN_4322 GBV_ILN_4323 GBV_ILN_4324 GBV_ILN_4325 GBV_ILN_4335 GBV_ILN_4338 GBV_ILN_4367 GBV_ILN_4700 AR 8 2013 3, p e58744
language	English
source	In PLoS ONE 8(2013), 3, p e58744 volume:8 year:2013 number:3, p e58744
sourceStr	In PLoS ONE 8(2013), 3, p e58744 volume:8 year:2013 number:3, p e58744
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container_title	PLoS ONE
authorswithroles_txt_mv	Asif J Iqbal @@aut@@ Daniel Regan-Komito @@aut@@ Ivy Christou @@aut@@ Gemma E White @@aut@@ Eileen McNeill @@aut@@ Amy Kenyon @@aut@@ Lewis Taylor @@aut@@ Theodore S Kapellos @@aut@@ Edward A Fisher @@aut@@ Keith M Channon @@aut@@ David R Greaves @@aut@@
publishDateDaySort_date	2013-01-01T00:00:00Z
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author	Asif J Iqbal
spellingShingle	Asif J Iqbal misc Medicine misc R misc Science misc Q A real time chemotaxis assay unveils unique migratory profiles amongst different primary murine macrophages.
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topic_title	A real time chemotaxis assay unveils unique migratory profiles amongst different primary murine macrophages
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title	A real time chemotaxis assay unveils unique migratory profiles amongst different primary murine macrophages.
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title_full	A real time chemotaxis assay unveils unique migratory profiles amongst different primary murine macrophages
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author_browse	Asif J Iqbal Daniel Regan-Komito Ivy Christou Gemma E White Eileen McNeill Amy Kenyon Lewis Taylor Theodore S Kapellos Edward A Fisher Keith M Channon David R Greaves
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title_sort	real time chemotaxis assay unveils unique migratory profiles amongst different primary murine macrophages
title_auth	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.
abstractGer	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.
abstract_unstemmed	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.
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title_short	A real time chemotaxis assay unveils unique migratory profiles amongst different primary murine macrophages.
url	https://doi.org/10.1371/journal.pone.0058744 https://doaj.org/article/fce6674479494768832c2d37a8acafe2 http://europepmc.org/articles/PMC3597586?pdf=render https://doaj.org/toc/1932-6203
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A real time chemotaxis assay unveils unique migratory profiles amongst different primary murine macrophages.

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