Wound Assay to Evaluate the Migratory Capacity of Murine Myoblasts in Co-Culture

In response to a wound, secretory cells produce paracrine signaling molecules to activate myoblasts — undifferentiated muscle cells. Activated myoblasts migrate toward the wound site, where they proliferate, causing the wound to heal.

To evaluate the migratory capacity of myoblasts in vitro, take a multiwell plate containing a confluent monolayer of murine myoblasts. Using a pipette tip, scratch the monolayer to create a linear, cell-free patch inside the well, which simulates the wound site.

Carefully supplement the well with the growth medium without disturbing the wound patch.

Take a membranous insert containing an adherent culture of neural crest cells, or NCCs — secretory cells — in a suitable medium. Place the insert inside the well, so that the NCCs at the base are near the wound patch. Incubate the co-culture for the desired duration.

During incubation, the NCCs release signaling molecules that bind to the specific receptors on the myoblasts, initiating a signaling cascade. This signaling facilitates actin reorganization in the myoblasts, and as a result, these cells develop cytoplasmic projections such as lamellipodia and filopodia, at their edges. These projections help t