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Advanced 2D Wound Healing Assay Procedures Performed using a Novel, Automated Scratch Tool, High Contrast Brightfield and Fluorescence Kinetic ImagingDownload
February 06, 2019
Author: Brad Larson, BioTek Instruments, Inc., Winooski, VT, USA
The ECM is a complex cellular scaffold found throughout the body that consists of collagen, cell-adhesive glycoproteins and other proteins. ECM supports adhesion and intra-cellular communication networks, and also facilitates cellular migration, including directionality, as a collective group. This type of migration is a significant aspect of wound healing and also of tumor metastasis, which is the primary cause of cancer morbidity and mortality. Additionally, stromal cells, such as fibroblasts, are responsible for depositing components of the ECM, and in the solid tumor microenvironment, influence cancer cells in migration, invasion and other tumorigenic processes. Because of the important role that ECM and stroma play in vivo, each should be included when performing in vitro cell migration studies to increase the relevance of generated data.
Here we demonstrate the use of a novel, automated tool to create consistent and reproducible scratch wounds in 2D cell monolayers formed on the bottom of a microplate. We use a single cancer cell model as well as a co-culture of fibroblasts and cancer cells; each plated in collagen-coated microplate wells to more closely simulate the in vivo tumor microenvironment and facilitate cell migration. The tool fits into any size laminar flow hood and features an interchangeable pin manifold for use with 24- or 96-well microplates, as well as a pre-programmed, hands-free cleaning and decontamination protocol to reduce the risk of buildup and contamination. Following scratch wound generation, the plate is transferred to BioTek’s automated imagers or the BioSpa™ Live Cell Imaging System to kinetically monitor cell migration using high contrast brightfield and fluorescence imaging.