Fig. 6. A proof-of-concept wound-healing assay using one dish precoated with Matrigel and fibronectin in different regions.
(A) Cartoon illustrating workflow. (a) A thin layer of medium is overlaid with FC40. (b) Two chambers (3 mm × 4 mm each) are printed side by side. (c) Surfaces in chambers are coated with Matrigel or fibronectin (2 μl; final concentration of 1 μg/cm2; 1 hour); the inset shows an image of one chamber. Fluid walls are now destroyed, and the dish is washed with 3 ml of medium to remove unattached coatings. (d) HEK cells (600,000) are plated in the dish. (e) After 24 hours, cells have formed a monolayer, and a wound (0.4 mm × 2 mm) is created by scraping the stylus over the surface to remove cells in its path. Healing of the wound is now monitored microscopically. (B and C) Images of wounds in monolayers grown on Matrigel or fibronectin. (a and b) Immediately before and after wounding (some droplets of FC40 remain where walls originally stood). (c) After 24 hours, cell growth reduces wound widths to <0.2 mm and <0.15 mm with Matrigel and fibronectin, respectively. (d) By 48 hours, wounds have completely healed.