Figure 2. Procedure to generate multiple wounds using a 3D-printed punch template.

(a) Circular wounds of 1 mm in diameter were made using a punch template in areas equidistant to the center of the culture. (b–d) The medium was removed before inserting the template into the culture well. Wounds were produced by pressing and rotating the metal cutting edge of the punch. Cells within the wound were debrided by rotation of a 10 μl disposable tip connected to a cordless precision screwdriver. (e–f) The circular wounds can be observed after staining with crystal violet.