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Agarose Spot as a Comparative Method for in situ Analysis of Simultaneous Chemotactic Responses to Multiple Chemokines.
Ahmed, Mohaned; Basheer, Haneen A; Ayuso, Jose M; Ahmet, Djevdet; Mazzini, Marco; Patel, Roshan; Shnyder, Steven D; Vinader, Victoria; Afarinkia, Kamyar.
Afiliação
  • Ahmed M; The Institute of Cancer Therapeutics, University of Bradford, West Yorkshire, BD7 1DP, United Kingdom.
  • Basheer HA; The Institute of Cancer Therapeutics, University of Bradford, West Yorkshire, BD7 1DP, United Kingdom.
  • Ayuso JM; Group of Structural Mechanics and Material Modelling, Universidad Zaragoza, Zaragoza, Spain.
  • Ahmet D; Department of Biomedical Engineering, Wisconsin Institutes for Medical Research, and The University of Wisconsin Carbone Cancer Center Madison, University of Wisconsin-Madison, Madison, WI 53715, USA.
  • Mazzini M; The Institute of Cancer Therapeutics, University of Bradford, West Yorkshire, BD7 1DP, United Kingdom.
  • Patel R; Dipartimento di Scienza e Tecnologia del Farmaco, Universitá Degli Studi di Torino, Via P. Giuria 9, 10125, Torino, Italy.
  • Shnyder SD; The Institute of Cancer Therapeutics, University of Bradford, West Yorkshire, BD7 1DP, United Kingdom.
  • Vinader V; The Institute of Cancer Therapeutics, University of Bradford, West Yorkshire, BD7 1DP, United Kingdom.
  • Afarinkia K; The Institute of Cancer Therapeutics, University of Bradford, West Yorkshire, BD7 1DP, United Kingdom.
Sci Rep ; 7(1): 1075, 2017 04 21.
Article em En | MEDLINE | ID: mdl-28432337
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.
Assuntos

Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: Quimiotaxia / Técnicas Citológicas / Quimiocinas / Células Epiteliais Limite: Humans Idioma: En Ano de publicação: 2017 Tipo de documento: Article

Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: Quimiotaxia / Técnicas Citológicas / Quimiocinas / Células Epiteliais Limite: Humans Idioma: En Ano de publicação: 2017 Tipo de documento: Article