The aim of this work is to show a novel method to evaluate the ability of some immunomodulatory molecules, such as antimicrobial peptides (AMPs), to stimulate cell migration. Importantly, cell migration is a rate-limiting event during the wound-healing process to re-establish the integrity and normal function of tissue layers after injury. The advantage of this method over the classical assay, which is based on a manually made scratch in a cell monolayer, is the usage of special silicone culture inserts providing two compartments to create a cell-free pseudo-wound field with a well-defined width (500 μm). In addition, due to an automated image analysis platform, it is possible to rapidly obtain quantitative data on the speed of wound closure and cell migration. More precisely, the effect of two frog-skin AMPs on the migration of bronchial epithelial cells will be shown. Furthermore, pretreatment of these cells with specific inhibitors will provide information on the molecular mechanisms underlying such events.

A novel in vitro wound healing assay to evaluate cell migration / Cappiello, Floriana; Casciaro, Bruno; Mangoni, Maria Luisa. - In: JOURNAL OF VISUALIZED EXPERIMENTS. - ISSN 1940-087X. - STAMPA. - 2018:133(2018). [10.3791/56825]

A novel in vitro wound healing assay to evaluate cell migration

Cappiello, Floriana;Casciaro, Bruno;Mangoni, Maria Luisa
2018

Abstract

The aim of this work is to show a novel method to evaluate the ability of some immunomodulatory molecules, such as antimicrobial peptides (AMPs), to stimulate cell migration. Importantly, cell migration is a rate-limiting event during the wound-healing process to re-establish the integrity and normal function of tissue layers after injury. The advantage of this method over the classical assay, which is based on a manually made scratch in a cell monolayer, is the usage of special silicone culture inserts providing two compartments to create a cell-free pseudo-wound field with a well-defined width (500 μm). In addition, due to an automated image analysis platform, it is possible to rapidly obtain quantitative data on the speed of wound closure and cell migration. More precisely, the effect of two frog-skin AMPs on the migration of bronchial epithelial cells will be shown. Furthermore, pretreatment of these cells with specific inhibitors will provide information on the molecular mechanisms underlying such events.
2018
amphibian skin; antimicrobial peptides; bronchial cells; cell migration; epidermal growth factor receptor; immunology and infection; innate immunity; issue 133; wound healing; neuroscience (all); chemical engineering (all); biochemistry, genetics and molecular biology (all); immunology and microbiology (all)
01 Pubblicazione su rivista::01a Articolo in rivista
A novel in vitro wound healing assay to evaluate cell migration / Cappiello, Floriana; Casciaro, Bruno; Mangoni, Maria Luisa. - In: JOURNAL OF VISUALIZED EXPERIMENTS. - ISSN 1940-087X. - STAMPA. - 2018:133(2018). [10.3791/56825]
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Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/11573/1135587
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