Optimization of the Wound Scratch Assay to Detect Changes in Murine Mesenchymal Stromal Cell Migration After Damage by Soluble Cigarette Smoke Extract

Abstract

Cell migration is vital to many physiological and pathological processes including tissue development, repair, and regeneration, cancer metastasis, and inflammatory responses. Given the current interest in the role of mesenchymal stromal cells in mediating tissue repair, we are interested in quantifying the migratory capacity of these cells, and understanding how migratory capacity may be altered after damage. Optimization of a rigorously quantitative migration assay that is both easy to customize and cost-effective to perform is key to answering questions concerning alterations in cell migration in response to various stimuli. Current methods for quantifying cell migration, including scratch assays, trans-well migration assays (Boyden chambers), micropillar arrays, and cell exclusion zone assays, possess a range of limitations in reproducibility, customizability, quantification, and cost-effectiveness. Despite its prominent use, the scratch assay is confounded by issues with reproducibility related to damage of the cell microenvironment, impediments to cell migration, influence of neighboring senescent cells, and cell proliferation, as well as lack of rigorous quantification. The optimized scratch assay described here demonstrates robust outcomes, quantifiable and image-based analysis capabilities, cost-effectiveness, and adaptability to other applications. Video LinkThe video component of this article can be found at

Stimulation of glycolysis promotes cardiomyocyte proliferation after injury in adult zebrafish

“…CMs were cultured in growth medium until the plates were confluent and then transfected with adenovirus vectors encoding the gene of interest. After 24 h of culture, a sterilized plastic ruler was laid on the plate, and a sterile pipette tip was used to make a scratch in the CM layer as previously described (Cormier et al, 2015). After removing the dead cells, the plates were incubated for 3 days and then analyzed.…”

Inhibiting endoplasmic reticulum stress decreases tumor burden in a mouse model for hepatocellular carcinoma

“…We also observed a direct effect of 4μ8C on LX2 and HepG2-migration, since treatment with 4μ8C lead to a significant reduction in wound closure after 24h, compared to untreated controls. It is important to note that traditional scratch wound assays cannot distinguish between proliferation and migration 13 . Considering that we observed a direct effect of 4μ8C on cell proliferation in HepG2-monocultures, this could have interfered with the quantification of wound closure.…”

Assembly of Fully Substituted 2H‐Indazoles Catalyzed by Cu2O Rhombic Dodecahedra and Evaluation of Anticancer Activity

“…Analysis of the effect of compound 3 n on migration of MDA‐MB‐468, was performed by using a wound healing assay . Cells were seeded at a density of 3×10 5 cells per well in a six‐well plate and incubated for 24 h. A single wound was made on the plates by using a 200 μL sterile pipette tip.…”