scholarly journals A Novel In Vitro Wound Healing Assay to Evaluate Cell Migration

10.3791/56825 ◽  
2018 ◽  
Author(s):  
Floriana Cappiello ◽  
Bruno Casciaro ◽  
Maria Luisa Mangoni
Keyword(s):  
Wound Healing ◽  
Cell Migration ◽  
Wound Healing Assay

A Method for Quantitative Analysis of the Kinematics of Fibroblast Migration in a Monolayer Wound Model

2011 ◽  
Author(s):  
Gil Topman ◽  
Orna Sharabani-Yosef ◽  
Amit Gefen
Keyword(s):  
Wound Healing ◽  
Cell Migration ◽  
Wound Healing Assay ◽  
Complete Coverage ◽  
Time Intervals ◽  
Fibroblast Migration ◽  
Wound Model ◽  
Regular Time

A wound healing assay is simple but effective method to study cell migration in vitro. Cell migration in vitro was found to mimic migration in vivo to some extent [1,2]. In wound healing assays, a “wound” is created by either scraping or mechanically crushing cells in a monolayer, thereby forming a denuded area. Cells migrate into the denuded area to complete coverage, and thereby “heal” the wound. Micrographs at regular time intervals are captured during such experiments for analysis of the process of migration.

scholarly journals Anti-cancer effect of dihydrokaempferol in human malignant melanoma cell is mediated via inhibition of cell migration and invasion and up-regulation of NF-kB/MAPK signalling pathways

2016 ◽  
Vol 11 (4) ◽  
pp. 810
Author(s):  
Ning Zeng ◽  
Hong Qiu ◽  
Min Wu ◽  
Yi Xu ◽  
Hai-Ping Wang ◽  
...  
Keyword(s):  
Wound Healing ◽  
Cell Migration ◽  
Malignant Melanoma ◽  
Signalling Pathways ◽  
Migration And Invasion ◽  
Wound Healing Assay ◽  
Human Malignant Melanoma ◽  
Cell Migration And Invasion ◽  
Mapk Signalling

<p class="Abstract">The purpose of the present research work was to demonstrate the antitumor activity of dihydrokaempferol in SK-Mel-28 human malignant melanoma cells. MTT assay was used to study the cytotoxic effects induced by dihydrokaempferol in these cells. In vitro wound healing assay and invasion assay were used to examine its effects on cell migration and invasion. Fluorescence microscopy using acridine orange/propidium iodide was used to study effects on cell morphology and apoptosis. Western blot assay revealed its effects on NF-kB/mitogen-activated protein kinase (MAPK) protein expression levels. The results indicated that dihydrokaempferol significantly inhibited the growth of these cells and the cytotoxicity pattern was shown to follow the drug dose and incubation times. Dihydrokaempferol led to onset of red fluorescence in these cells indicating that its treatment with different doses leads to induction of apoptosis. Dihydrokaempferol also led to inhibition of cell migration and invasion in a dose-dependent manner. It was also shown to up-regulate NF-kB/MAPK signalling pathways.</p><p class="Abstract"><strong>Video Clip:</strong></p><p class="Abstract"><a href="https://youtube.com/v/g8vkXiPHG4A"><em>In vitro</em> wound healing assay:</a> 4 min 25 sec</p><p> </p>

Wound Healing Assay for Melanoma Cell Migration

2021 ◽  
pp. 65-71
Author(s):  
Juliano T. Freitas ◽  
Ivan Jozic ◽  
Barbara Bedogni
Keyword(s):  
Wound Healing ◽  
Cell Migration ◽  
Melanoma Cell ◽  
Wound Healing Assay

scholarly journals Role of Berry Anthocyanins and Phenolic Acids on Cell Migration and Angiogenesis: An Updated Overview

Nutrients ◽  
2019 ◽  
Vol 11 (5) ◽  
pp. 1075 ◽  
Author(s):  
Panagiotis Tsakiroglou ◽  
Natalie E. VandenAkker ◽  
Cristian Del Bo’ ◽  
Patrizia Riso ◽  
Dorothy Klimis-Zacas
Keyword(s):  
Wound Healing ◽  
Cell Migration ◽  
Phenolic Acids ◽  
Rho Gtpase ◽  
Small Gtpases ◽  
Endothelial Cell Migration ◽  
Berry Extracts

Cell migration is a critical process that is highly involved with normal and pathological conditions such as angiogenesis and wound healing. Important members of the RHO GTPase family are capable of controlling cytoskeleton conformation and altering motility characteristics of cells. There is a well-known relationship between small GTPases and the PI3K/AKT pathway. Endothelial cell migration can lead to angiogenesis, which is highly linked to wound healing processes. Phenolics, flavonoids, and anthocyanins are major groups of phytochemicals and are abundant in many natural products. Their antioxidant, antimicrobial, anti-inflammatory, antidiabetic, angiogenenic, neuroprotective, hepatoprotective, and cardioprotective properties have been extensively documented. This comprehensive review focuses on the in vitro and in vivo role of berry extracts and single anthocyanin and phenolic acid compounds on cell migration and angiogenesis. We aim to summarize the most recent published studies focusing on the experimental model, type of berry extract, source, dose/concentration and overall effect(s) of berry extracts, anthocyanins, and phenolic acids on the above processes.

scholarly journals Extracellular Signal-Regulated Kinase/Mitogen-Activated Protein Kinase Regulates Actin Organization and Cell Motility by Phosphorylating the Actin Cross-Linking Protein EPLIN

2007 ◽  
Vol 27 (23) ◽  
pp. 8190-8204 ◽  
Author(s):  
Mei-Ying Han ◽  
Hidetaka Kosako ◽  
Toshiki Watanabe ◽  
Seisuke Hattori
Keyword(s):  
Wound Healing ◽  
Cell Migration ◽  
Cell Motility ◽  
Stress Fiber ◽  
Mitogen Activated Protein Kinase ◽  
Cross Linking ◽  
Intact Cells ◽  
Extracellular Signal Regulated Kinase ◽  
Extracellular Signal

ABSTRACT Extracellular signal-regulated kinase (ERK) is important for various cellular processes, including cell migration. However, the detailed molecular mechanism by which ERK promotes cell motility remains elusive. Here we characterize epithelial protein lost in neoplasm (EPLIN), an F-actin cross-linking protein, as a novel substrate for ERK. ERK phosphorylates Ser360, Ser602, and Ser692 on EPLIN in vitro and in intact cells. Phosphorylation of the C-terminal region of EPLIN reduces its affinity for actin filaments. EPLIN colocalizes with actin stress fibers in quiescent cells, and stimulation with platelet-derived growth factor (PDGF) induces stress fiber disassembly and relocalization of EPLIN to peripheral and dorsal ruffles, wherein phosphorylation of Ser360 and Ser602 is observed. Phosphorylation of these two residues is also evident during wound healing at the leading edge of migrating cells. Moreover, expression of a non-ERK-phosphorylatable mutant, but not wild-type EPLIN, prevents PDGF-induced stress fiber disassembly and membrane ruffling and also inhibits wound healing and PDGF-induced cell migration. We propose that ERK-mediated phosphorylation of EPLIN contributes to actin filament reorganization and enhanced cell motility.

IL-6/STAT3/TFF3 signaling regulates human biliary epithelial cell migration and wound healing in vitro

2010 ◽  
Vol 37 (8) ◽  
pp. 3813-3818 ◽  
Author(s):  
Gui-xing Jiang ◽  
Xiang-yu Zhong ◽  
Yun-fu Cui ◽  
Wei Liu ◽  
Sheng Tai ◽  
...  
Keyword(s):  
Wound Healing ◽  
Cell Migration ◽  
Epithelial Cell ◽  
Biliary Epithelial Cell ◽  
Epithelial Cell Migration

scholarly journals An image J plugin for the high throughput image analysis of in vitro scratch wound healing assays

2020 ◽  
Author(s):  
Alejandra Suarez-Arnedo ◽  
Felipe Torres Figueroa ◽  
Camila Clavijo ◽  
Pablo Arbeláez ◽  
Juan C. Cruz ◽  
...  
Keyword(s):  
Wound Healing ◽  
Image Analysis ◽  
Low Cost ◽  
Cellular Migration ◽  
Wound Healing Assay ◽  
Wound Area ◽  
Scratch Wound ◽  
Scratch Wound Healing Assay ◽  
Imagej Plugin

AbstractIn vitro scratch wound healing assay, a simple and low-cost technique that works along with other image analysis tools, is one of the most widely used 2D methods to determine the cellular migration and proliferation in processes such as regeneration and disease. There are open-source programs such as imageJ to analyze images of in vitro scratch wound healing assays, but these tools require manual tuning of various parameters, which is time-consuming and limits image throughput. For that reason, we developed an optimized plugin for imageJ to automatically recognize the wound healing size, correct the average wound width by considering its inclination, and quantify other important parameters such as: area, wound area fraction, average wound width, and width deviation of the wound images obtained from a scratch/ wound healing assay. Our plugin is easy to install and can be used with different operating systems. It can be adapted to analyze both individual images and stacks. Additionally, it allows the analysis of images obtained from bright field, phase contrast, and fluorescence microscopes. In conclusion, this new imageJ plugin is a robust tool to automatically standardize and facilitate quantification of different in vitro wound parameters with high accuracy compared with other tools and manual identification.

scholarly journals Activator Protein-1 Transcriptional Activity Drives Soluble Micrograft-Mediated Cell Migration and Promotes the Matrix Remodeling Machinery

2019 ◽  
Vol 2019 ◽  
pp. 1-19 ◽  
Author(s):  
Martina Balli ◽  
Jonathan Sai-Hong Chui ◽  
Paraskevi Athanasouli ◽  
Willy Antoni Abreu de Oliveira ◽  
Youssef El Laithy ◽  
...  
Keyword(s):  
Wound Healing ◽  
Cell Migration ◽  
Wound Repair ◽  
Molecular Mechanisms ◽  
Cell Model ◽  
Matrix Remodeling ◽  
Activator Protein 1 ◽  
Impaired Wound Healing ◽  
Beneficial Effects

Impaired wound healing and tissue regeneration have severe consequences on the patient’s quality of life. Micrograft therapies are emerging as promising and affordable alternatives to improve skin regeneration by enhancing the endogenous wound repair processes. However, the molecular mechanisms underpinning the beneficial effects of the micrograft treatments remain largely unknown. In this study, we identified the active protein-1 (AP-1) member Fos-related antigen-1 (Fra-1) to play a central role in the extracellular signal-regulated kinase- (ERK-) mediated enhanced cell migratory capacity of soluble micrograft-treated mouse adult fibroblasts and in the human keratinocyte cell model. Accordingly, we show that increased micrograft-dependent in vitro cell migration and matrix metalloprotease activity is abolished upon inhibition of AP-1. Furthermore, soluble micrograft treatment leads to increased expression and posttranslational phosphorylation of Fra-1 and c-Jun, resulting in the upregulation of wound healing-associated genes mainly involved in the regulation of cell migration. Collectively, our work provides insights into the molecular mechanisms behind the cell-free micrograft treatment, which might contribute to future advances in wound repair therapies.

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