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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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    Publication date
    2017-04-21
    Author
    Ahmed, Mohaned S.A.
    Basheer, Haneen A.
    Ayuso, J.M.
    Ahmet, Djevdet S.
    Mazzini, Marco
    Patel, Roshan
    Shnyder, Steven D.
    Vinader, Victoria
    Afarinkia, Kamyar
    Keyword
    Cancer models; Drug screening; Chemotactic migration; Agarose gel; Chemokines
    Rights
    © 2017 The Authors. Published by Nature Publishing. Open Access. This article is licensed under a Creative Commons Attribution 4.0 International License.
    Peer-Reviewed
    yes
    
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    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.
    URI
    http://hdl.handle.net/10454/11904
    Version
    Published version
    Citation
    Ahmed M, Basheer HA, Ayuso JM, Ahmet D, Mazzini M, Patel R, Shnyder SD, Vinader V and Afarinkia K (2017) Agarose Spot as a Comparative Method for in situ Analysis of Simultaneous Chemotactic Responses to Multiple Chemokines. Scientific Reports. 7: 1075.
    Link to publisher’s version
    https://doi.org/10.1038/s41598-017-00949-4
    Type
    Article
    Collections
    Life Sciences Publications

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