Advances in Wound-Healing Assays for Probing Collective Cell Migration

Department of Aerospace and Mechanical Engineering, The University of Arizona, Tucson, AZ, USA.
Journal of the Association for Laboratory Automation (Impact Factor: 1.5). 02/2012; 17(1):59-65. DOI: 10.1177/2211068211426550
Source: PubMed


Collective cell migration plays essential roles in a wide spectrum of biological processes, such as embryogenesis, tissue regeneration, and cancer metastasis. Numerous wound-healing assays based on mechanical, chemical, optical, and electrical approaches have been developed to create model "wounds" in cell monolayers to study the collective cell migration processes. These approaches can result in different microenvironments for cells to migrate and possess diverse assay characteristics in terms of simplicity, throughput, reproducibility, and multiplexability. In this review, we provide an overview of advances in wound-healing assays and discuss their advantages and limitations in studying collective cell migration.

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    • "Compared with simple proliferation assays, migration assays add another dimension and allow easy screening of many compounds. Wound healing assays, where confluent monolayers of ECs are mechanically or chemically wounded in a standardised manner, combine proliferation and migration and also allow easy screening end evaluation of the effects of many compounds [28]. "
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    ABSTRACT: Angiogenesis, the formation of new blood vessels from existing vessels is required for many physiological processes and for growth of solid tumors. Initiated by hypoxia, angiogenesis involves binding of angiogenic factors to endothelial cell (EC) receptors and activation of cellular signaling, differentiation, migration, proliferation, interconnection and canalization of ECs, remodeling of the extracellular matrix and stabilization of newly formed vessels. Experimentally, these processes can be studied by several in vitro and in vivo assays focusing on different steps in the process. In vitro, ECs form networks of capillary-like tubes when propagated for three days in coculture with fibroblasts. The tube formation is dependent on vascular endothelial growth factor (VEGF) and omission of VEGF from the culture medium results in the formation of clusters of undifferentiated ECs. Addition of angiogenesis inhibitors to the coculture system disrupts endothelial network formation and influences EC morphology in two distinct ways. Treatment with antibodies to VEGF, soluble VEGF receptor, the VEGF receptor tyrosine kinase inhibitor SU5614, protein tyrosine phosphatase inhibitor (PTPI) IV or levamisole results in the formation of EC clusters of variable size. This cluster morphology is a result of inhibited EC differentiation and levamisole can be inferred to influence and block VEGF signaling. Treatment with platelet factor 4, thrombospondin, rapamycin, suramin, TNP-470, salubrinal, PTPI I, PTPI II, clodronate, NSC87877 or non-steriodal anti-inflammatory drugs (NSAIDs) results in the formation of short cords of ECs, which suggests that these inhibitors have an influence on later steps in the angiogenic process, such as EC proliferation and migration. A humanized antibody to VEGF is one of a few angiogenesis inhibitors used clinically for treatment of cancer. Levamisole is approved for clinical treatment of cancer and is interesting with respect to anti-angiogenic activity in vivo since it inhibits ECs in vitro with a morphology resembling that obtained with antibodies to VEGF.
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    • "In these assays, a cell layer is allowed to move into a cell-free region, and the decrease in time of this area reflects cell migration velocity. The cell-free zone can be created (Riahi et al., 2012) by scratching a confluent cell layer [typical wound healing assay (Liang et al., 2007)] or by using non-damaging barriers during cell seeding [cell exclusion zone assay (Gough et al., 2011)]. Advanced microscopes then allow automatic acquisition of images of the migrating cells, and specialized image processing tools are used to quantify cell migration features from these images (Table 1). "
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    ABSTRACT: Automated image processing has allowed cell migration research to evolve to a high-throughput research field. As a consequence, there is now an unmet need for data management in this domain. The absence of a generic management system for the quantitative data generated in cell migration assays results in each dataset being treated in isolation, making data comparison across experiments difficult. Moreover, by integrating quality control and analysis capabilities into such a data management system, the common practice of having to manually transfer data across different downstream analysis tools will be markedly sped up and made more robust. In addition, access to a data management solution creates gateways for data standardization, meta-analysis and structured public data dissemination. We here present CellMissy, a cross-platform data management system for cell migration data with a focus on wound healing data. CellMissy simplifies and automates data management, storage and analysis from the initial experimental set-up to data exploration. Availability and implementation: CellMissy is a cross-platform open-source software developed in Java. Source code and cross-platform binaries are freely available under the Apache2 open source license at Contact: Supplementary Information: Supplementary data are available at Bioinformatics online.
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