![]() ![]() The CWCA uses a sterile 10 μl (P10) micropipette tip attached to an aspirator to remove a small circular area of cells ( Figure 1). One advantage to these techniques is that they can generate highly consistent initial wounds however, they are more complex and costlier than the CWCA described here. Current techniques to generate circular wounds, such as exclusion zone assays, involve growing the cells around circular barriers (poly-dimethylsiloxane micropillars, stoppers, or biocompatible gels) of uniform size, or using a stabilized, rotating, silicone-tipped drill press to create uniform, circular wounds in an intact confluent monolayer of cells ( Table 1). The circular wound closure assay (CWCA) permits the analyst to easily relocate the wound at any time point, and it enables accurate analysis by calculating the area or the radius of the circular wound using image analysis software. The concept for generating a circular wound for measuring 2D cell migration has been previously established. Gel needs to be manually removed, which may alter cell or substrate properties.(1) Apply gel in the center of each well prior to seeding cells Unknown effects of stopper-derived components on cell propertiesĬell exclusion zone assay with biocompatible gel.(3) Remove stopper to expose circular wound (2) Allow cells to grow around the stopper ![]() (1) Insert stopper in well prior to seeding cells
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