A Haptotaxis Assay for Neutrophils using Optical Patterning and a High-content Approach

Abstract Neutrophil recruitment guided by chemotactic cues is a central event in host defense against infection and tissue injury. While the mechanisms underlying neutrophil chemotaxis have been extensively studied, these are just recently being addressed by using high-content approaches or surface-...

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Autores principales:	Joannie Roy, Javier Mazzaferri, János G. Filep, Santiago Costantino
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2017
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/d693fc036fee42848cb71caa988ac6fc
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spelling	oai:doaj.org-article:d693fc036fee42848cb71caa988ac6fc2021-12-02T16:06:52ZA Haptotaxis Assay for Neutrophils using Optical Patterning and a High-content Approach10.1038/s41598-017-02993-62045-2322https://doaj.org/article/d693fc036fee42848cb71caa988ac6fc2017-06-01T00:00:00Zhttps://doi.org/10.1038/s41598-017-02993-6https://doaj.org/toc/2045-2322Abstract Neutrophil recruitment guided by chemotactic cues is a central event in host defense against infection and tissue injury. While the mechanisms underlying neutrophil chemotaxis have been extensively studied, these are just recently being addressed by using high-content approaches or surface-bound chemotactic gradients (haptotaxis) in vitro. Here, we report a haptotaxis assay, based on the classic under-agarose assay, which combines an optical patterning technique to generate surface-bound formyl peptide gradients as well as an automated imaging and analysis of a large number of migration trajectories. We show that human neutrophils migrate on covalently-bound formyl-peptide gradients, which influence the speed and frequency of neutrophil penetration under the agarose. Analysis revealed that neutrophils migrating on surface-bound patterns accumulate in the region of the highest peptide concentration, thereby mimicking in vivo events. We propose the use of a chemotactic precision index, gyration tensors and neutrophil penetration rate for characterizing haptotaxis. This high-content assay provides a simple approach that can be applied for studying molecular mechanisms underlying haptotaxis on user-defined gradient shape.Joannie RoyJavier MazzaferriJános G. FilepSantiago CostantinoNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 7, Iss 1, Pp 1-13 (2017)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Joannie Roy Javier Mazzaferri János G. Filep Santiago Costantino A Haptotaxis Assay for Neutrophils using Optical Patterning and a High-content Approach
description	Abstract Neutrophil recruitment guided by chemotactic cues is a central event in host defense against infection and tissue injury. While the mechanisms underlying neutrophil chemotaxis have been extensively studied, these are just recently being addressed by using high-content approaches or surface-bound chemotactic gradients (haptotaxis) in vitro. Here, we report a haptotaxis assay, based on the classic under-agarose assay, which combines an optical patterning technique to generate surface-bound formyl peptide gradients as well as an automated imaging and analysis of a large number of migration trajectories. We show that human neutrophils migrate on covalently-bound formyl-peptide gradients, which influence the speed and frequency of neutrophil penetration under the agarose. Analysis revealed that neutrophils migrating on surface-bound patterns accumulate in the region of the highest peptide concentration, thereby mimicking in vivo events. We propose the use of a chemotactic precision index, gyration tensors and neutrophil penetration rate for characterizing haptotaxis. This high-content assay provides a simple approach that can be applied for studying molecular mechanisms underlying haptotaxis on user-defined gradient shape.
format	article
author	Joannie Roy Javier Mazzaferri János G. Filep Santiago Costantino
author_facet	Joannie Roy Javier Mazzaferri János G. Filep Santiago Costantino
author_sort	Joannie Roy
title	A Haptotaxis Assay for Neutrophils using Optical Patterning and a High-content Approach
title_short	A Haptotaxis Assay for Neutrophils using Optical Patterning and a High-content Approach
title_full	A Haptotaxis Assay for Neutrophils using Optical Patterning and a High-content Approach
title_fullStr	A Haptotaxis Assay for Neutrophils using Optical Patterning and a High-content Approach
title_full_unstemmed	A Haptotaxis Assay for Neutrophils using Optical Patterning and a High-content Approach
title_sort	haptotaxis assay for neutrophils using optical patterning and a high-content approach
publisher	Nature Portfolio
publishDate	2017
url	https://doaj.org/article/d693fc036fee42848cb71caa988ac6fc
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A Haptotaxis Assay for Neutrophils using Optical Patterning and a High-content Approach

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