A novel approach to quantify the wound closure dynamic

• Published in: Experimental cell research Vol. 352; no. 2; pp. 175 - 183
• Main Authors: Ascione, Flora, Guarino, Andrea Maria, Calabrò, Viola, Guido, Stefano, Caserta, Sergio
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 15.03.2017
• Subjects: Breast Cancer; Cell Line, Tumor; Cell Migration; Cell Migration Assays - instrumentation; Cell Migration Assays - methods; Cell Movement; Cell Proliferation; Diffusion; Humans; Image Analysis; Mathematical modeling; Microscopy, Fluorescence - instrumentation; Microscopy, Fluorescence - methods; Re-Epithelialization; Time-lapse Microscopy; Transport Phenomena; Wound Healing; Cell Migration; Transport Phenomena; Time-lapse Microscopy; Cell Proliferation; Mathematical modeling; Wound Healing; Diffusion; Image Analysis; Breast Cancer
• ISSN: 0014-4827; 1090-2422
• DOI: 10.1016/j.yexcr.2017.01.005

The Wound Healing (WH) assay is widely used to investigate cell migration in vitro, in order to reach a better understanding of many physiological and pathological phenomena. Several experimental factors, such as uneven cell density among different samples, can affect the reproducibility and reliability of this assay, leading to a discrepancy in the wound closure kinetics among data sets corresponding to the same cell sample. We observed a linear relationship between the wound closure velocity and cell density, and suggested a novel methodological approach, based on transport phenomena concepts, to overcome this source of error on the analysis of the Wound Healing assay. In particular, we propose a simple scaling of the experimental data, based on the interpretation of the wound closure as a diffusion-reaction process. We applied our methodology to the MDA-MB-231 breast cancer cells, whose motility was perturbed by silencing or over-expressing genes involved in the control of cell migration. Our methodological approach leads to a significant improvement in the reproducibility and reliability in the in vitro WH assay. [Display omitted] •Cell density can affect the reproducibility and reliability of wound healing assays.•A linear relationship between the wound closure velocity and cell density is observed.•A novel methodology of analysis is proposed to account for cell density effect.•The motility of three populations of MDA-MB-231 breast cancer cells is compared.•YB-1 depleted cells show higher motility, ∆Np63α transfected cells have lower motility.