Agarose Spot as a Comparative Method for in situ Analysis of Simultaneous Chemotactic Responses to Multiple Chemokines

Abstract We describe a novel protocol to quantitatively and simultaneously compare the chemotactic responses of cells towards different chemokines. In this protocol, droplets of agarose gel containing different chemokines are applied onto the surface of a Petri dish, and then immersed under culture...

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Autores principales:	Mohaned Ahmed, Haneen A. Basheer, Jose M. Ayuso, Djevdet Ahmet, Marco Mazzini, Roshan Patel, Steven D. Shnyder, Victoria Vinader, Kamyar Afarinkia
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2017
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/696870599d204173b0f1ab997124d247
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spelling	oai:doaj.org-article:696870599d204173b0f1ab997124d2472021-12-02T11:53:05ZAgarose Spot as a Comparative Method for in situ Analysis of Simultaneous Chemotactic Responses to Multiple Chemokines10.1038/s41598-017-00949-42045-2322https://doaj.org/article/696870599d204173b0f1ab997124d2472017-04-01T00:00:00Zhttps://doi.org/10.1038/s41598-017-00949-4https://doaj.org/toc/2045-2322Abstract We describe a novel protocol to quantitatively and simultaneously compare the chemotactic responses of cells towards different chemokines. In this protocol, droplets of agarose gel containing different chemokines are applied onto the surface of a Petri dish, and then immersed under culture medium in which cells are suspended. As chemokine molecules diffuse away from the spot, a transient chemoattractant gradient is established across the spots. Cells expressing the corresponding cognate chemokine receptors migrate against this gradient by crawling under the agarose spots towards their centre. We show that this migration is chemokine-specific; meaning that only cells that express the cognate chemokine cell surface receptor, migrate under the spot containing its corresponding chemokine ligand. Furthermore, we show that migration under the agarose spot can be modulated by selective small molecule antagonists present in the cell culture medium.Mohaned AhmedHaneen A. BasheerJose M. AyusoDjevdet AhmetMarco MazziniRoshan PatelSteven D. ShnyderVictoria VinaderKamyar AfarinkiaNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 7, Iss 1, Pp 1-11 (2017)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Mohaned Ahmed Haneen A. Basheer Jose M. Ayuso Djevdet Ahmet Marco Mazzini Roshan Patel Steven D. Shnyder Victoria Vinader Kamyar Afarinkia Agarose Spot as a Comparative Method for in situ Analysis of Simultaneous Chemotactic Responses to Multiple Chemokines
description	Abstract We describe a novel protocol to quantitatively and simultaneously compare the chemotactic responses of cells towards different chemokines. In this protocol, droplets of agarose gel containing different chemokines are applied onto the surface of a Petri dish, and then immersed under culture medium in which cells are suspended. As chemokine molecules diffuse away from the spot, a transient chemoattractant gradient is established across the spots. Cells expressing the corresponding cognate chemokine receptors migrate against this gradient by crawling under the agarose spots towards their centre. We show that this migration is chemokine-specific; meaning that only cells that express the cognate chemokine cell surface receptor, migrate under the spot containing its corresponding chemokine ligand. Furthermore, we show that migration under the agarose spot can be modulated by selective small molecule antagonists present in the cell culture medium.
format	article
author	Mohaned Ahmed Haneen A. Basheer Jose M. Ayuso Djevdet Ahmet Marco Mazzini Roshan Patel Steven D. Shnyder Victoria Vinader Kamyar Afarinkia
author_facet	Mohaned Ahmed Haneen A. Basheer Jose M. Ayuso Djevdet Ahmet Marco Mazzini Roshan Patel Steven D. Shnyder Victoria Vinader Kamyar Afarinkia
author_sort	Mohaned Ahmed
title	Agarose Spot as a Comparative Method for in situ Analysis of Simultaneous Chemotactic Responses to Multiple Chemokines
title_short	Agarose Spot as a Comparative Method for in situ Analysis of Simultaneous Chemotactic Responses to Multiple Chemokines
title_full	Agarose Spot as a Comparative Method for in situ Analysis of Simultaneous Chemotactic Responses to Multiple Chemokines
title_fullStr	Agarose Spot as a Comparative Method for in situ Analysis of Simultaneous Chemotactic Responses to Multiple Chemokines
title_full_unstemmed	Agarose Spot as a Comparative Method for in situ Analysis of Simultaneous Chemotactic Responses to Multiple Chemokines
title_sort	agarose spot as a comparative method for in situ analysis of simultaneous chemotactic responses to multiple chemokines
publisher	Nature Portfolio
publishDate	2017
url	https://doaj.org/article/696870599d204173b0f1ab997124d247
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Agarose Spot as a Comparative Method for in situ Analysis of Simultaneous Chemotactic Responses to Multiple Chemokines

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