A Novel In Vitro Wound Healing Assay to Evaluate Cell Migration

被引:69
作者
Cappiello, Floriana [1 ]
Casciaro, Bruno [1 ]
Mangoni, Maria Luisa [1 ]
机构
[1] Sapienza Univ Rome, Dept Biochem Sci, Rome, Italy
来源
JOVE-JOURNAL OF VISUALIZED EXPERIMENTS | 2018年 / 133期
关键词
Immunology and Infection; Issue; 133; Bronchial Cells; Wound Healing; Cell Migration; Antimicrobial Peptides; Epidermal Growth Factor Receptor; Innate Immunity; Amphibian Skin; GROWTH-FACTOR RECEPTOR; ANTIMICROBIAL PEPTIDE LL-37; CYSTIC-FIBROSIS PATIENTS; AIRWAY; SKIN; TRANSACTIVATION; INFECTIONS; IMMUNITY; REPAIR; CFTR;
D O I
10.3791/56825
中图分类号
O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];
学科分类号
07 ; 0710 ; 09 ;
摘要
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 mu m). 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.
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页数:6
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