期刊
JOVE-JOURNAL OF VISUALIZED EXPERIMENTS
卷 -, 期 133, 页码 -出版社
JOURNAL OF VISUALIZED EXPERIMENTS
DOI: 10.3791/56825
关键词
Immunology and Infection; Issue 133; Bronchial Cells; Wound Healing; Cell Migration; Antimicrobial Peptides; Epidermal Growth Factor Receptor; Innate Immunity; Amphibian Skin
资金
- Sapienza University of Rome
- Italian Cystic Fibrosis Research Foundation [11/2014]
- FILAS [FILAS-RU-2014-1020]
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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