Research Techniques Made Simple: Analysis of Collective Cell Migration Using the Wound Healing Assay

Grada, Ayman; Otero-Viñas, Marta; Prieto‐Castrillo, Francisco; Obagi, Zaidal; Falanga, Vincent

doi:10.1016/j.jid.2016.11.020

Cited by 547 publications

(411 citation statements)

References 16 publications

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“…In addition, all results presented here were obtained from in vitro studies. Although the scratch assay is a simple, versatile, and cost-effective method, it has some limitations [44]. Thus, more complex methods, such as 3D cell culture systems and in vivo studies, are required to confirm our results.…”

Section: Discussionmentioning

confidence: 72%

Poly I:C induces collective migration of HaCaT keratinocytes via IL-8

et al. 2017

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BackgroundDelayed wound healing reduces the quality of life (QOL) of patients. Thus, understanding the mechanism of wound healing is indispensable for better management. However, the role of innate immunity in wound healing is thus far unknown. Recently the involvement of TLR3 in wound healing has been evaluated. The systemic administration of polyriboinosinic-polyribocytidylic acid (poly I:C ; a substitute for viral dsRNA and a ligand of toll-like receptor 3), enhances wound healing in vivo. The aim of this study is to improve our understanding of the link between innate immunity and human wound healing, particularly in re-epithelialization.ResultsThe present study showed that poly I:C significantly accelerated collective HaCaT cell migration in a scratch assay. Poly I:C also increased IL-8 and bFGF production, and anti-IL-8 antibodies significantly inhibited the migration caused by poly I:C. Human recombinant IL-8 also accelerated collective HaCaT cell migration. An immunofluorescence assay and enzyme-linked immunosorbent assay (ELISA) also revealed that poly I:C decreased E-cadherin protein levels and increased vimentin protein levels, and anti-IL-8 antibody reversed this effect. In contrast, nucleic/cytosolic protein ratios of Snail 1 were unchanged in all tested conditions.ConclusionOur findings demonstrated that poly I:C accelerated collective HaCaT cell migration via autocrine/paracrine secretions of IL-8 and the subsequent incomplete epithelial-mesenchymal transition (EMT). Our findings provide a new strategy for wound healing by regulating innate immune systems in re-epithelialization.

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Section: Discussionmentioning

confidence: 72%

Poly I:C induces collective migration of HaCaT keratinocytes via IL-8

et al. 2017

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“…Cell mobility could therefore be used as a parameter to assess the physiological state of cell. For that purpose scratch assay was performed, as it is a commonly used technique for assessment of the toxicity of the biocompound in dermatology . It consists to monitor wound healing ability of cells exposed to pyomelanin (100 μg/ml) as compared to nonexposed one (Figures and ).…”

Section: Resultsmentioning

confidence: 99%

Characterization of a nontoxic pyomelanin pigment produced by the yeast Yarrowia lipolytica

Tahar

Kus‐Liśkiewicz

Lara

et al. 2019

Biotechnology Progress

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In this study, we report on the ability of the yeast Yarrowia lipolytica W29 to produce an extracellular melanin‐like brown pigment at high yield (0.5 mg/ml) in culture medium supplemented with L‐tyrosine. This pigment has been characterized as pyomelanin and its synthesis was found to occur by the so‐called HGA‐melanin pathway. The purified pyomelanin was found embedded with antioxidant properties as it exhibited a radical scavenging activity toward 1,1‐diphenyl‐2‐picrylhydrazyl (DPPH) with IC50 of 230 μg/ml. It was also characterized as noncytotoxic toward two mammalian cell lines, namely the mouse fibroblast NIH3T3 and human keratinocytes HaCaT. When blended with different commercial sunscreens, the purified pyomelanin increased significantly the sun protection factor (SPF) value, highlighting its potential utilization as UV‐filter in cosmetic preparations.

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“…On the other hand, GRX detached from the substrate individually. These phenomena are very interesting because GRX EGFP‐Cav1 showed an enhanced mechanosensory, surface tension, and cell‐cell affinity to continually attached to others . Curiously, at time 6 hours, migration pattern of both cell lines is different.…”

Section: Resultsmentioning

confidence: 99%

“…These phenomena are very interesting because GRX EGFP-Cav1 showed an enhanced mechanosensory, surface tension, and cell-cell affinity to continually attached to others. 38 Curiously, at time 6 hours, migration pattern of both cell lines is different. GRX expanded pseudopodium-like projections into the free space and have a cell single migration.…”

Section: Cav-1 In Cell Migrationmentioning

confidence: 99%

Exogenous expression of caveolin‐1 is sufficient for hepatic stellate cell activation

Ilha

Moraes

Rohden

et al. 2019

J of Cellular Biochemistry

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Caveolin‐1 (Cav‐1) expression is increased in hepatic stellate cells (HSC) upon liver cirrhosis and it functions as an integral membrane protein of lipid rafts and caveolae that regulates and integrates multiple signals as a platform. This study aimed to evaluate the role of Cav‐1 in HSC. Thus, the effects of exogenous expression of Cav‐1 in GRX cells, a model of activated HSC, were determined. Here, we demonstrated through evaluating well‐known HSC activation markers – such as α‐smooth muscle actin, collagen I, and glial fibrillary acidic protein – that up regulation of Cav‐1 induced GRX to a more activated phenotype. GRXEGFP‐Cav1 presented an increased migration, an altered adhesion pattern, a reorganization f‐actin cytoskeleton, an arrested cell cycle, a modified cellular ultrastructure, and a raised endocytic flux. Based on this, GRX EGFP‐Cav1 represents a new cellular model that can be an important tool for understanding of events related to HSC activation. Furthermore, our results reinforce the role of Cav‐1 as a molecular marker of HSC activation.

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Research Techniques Made Simple: Analysis of Collective Cell Migration Using the Wound Healing Assay

Cited by 547 publications

References 16 publications

Poly I:C induces collective migration of HaCaT keratinocytes via IL-8

Poly I:C induces collective migration of HaCaT keratinocytes via IL-8

Characterization of a nontoxic pyomelanin pigment produced by the yeast Yarrowia lipolytica

Exogenous expression of caveolin‐1 is sufficient for hepatic stellate cell activation

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