Collective migration of an epithelial monolayer in response to a model wound
M. Poujade*, E. Grasland-Mongrain*, A. Hertzog†, J. Jouanneau†, P. Chavrier†, B. Ladoux‡, A. Buguin*, and P. Silberzan*§
As a result, the authors of this article orchestrated a very simple but elegant design to study the response of the monolayer as a result of a free surface. The experiment involved culturing a monolayer of cells constrained within a microstencil, which is fabricated from polydimethylsiloxane (PDMS) that precisely defined the geometrical conditions of the monolayer. The stencil was removed and the cells were free to migrate as shown in Figure 1. There are some advantages with this assay in observing cell migration. First, it eliminates the need to “scratch” the monolayer, which might depend on the user and the tool used to scratch the surface. Next, the wounds are well defined and perfectly controlled, allowing the authors to observe the cells progress from its initial position. The authors concluded that their injury-free wound healing assay on MDCK epithelial cells showed that the free surface was sufficient in inducing cell migration. Moreover, they note that the cells migrate in a leader-follower pattern which a certain cell will take “lead” in migration.
I chose this paper because of the simplicity of the experiment. For my project, I plan to use a similar design in determining cell migratory patterns within various defined regions.
7 comments:
As a result of the intracellular signaling left by the removed cells, do the remaining cells in the classical scratch assay also work through a follow-the-leader type migration, or is it more of a free for all?
Interesting question Alvin. It is still unclear whether the scratch process triggers the migration through the intracellular signaling or the motile response as a result of the free surface. However, I believe the cells would undergo the same migratory patterns in the classical scratch assay and the stencil assay. We might actually be doing our project in comparing this, so keep checking for more updates.
This leader-follower migration pattern sounds very interesting. Have the researchers identified any specific factors that make certain cells "leaders" and others not? Also, have they also tested whether the presence of leader cells necessary to induce a collective cell migration?
This is an interesting new way to observe the migration of cells. What geometric shapes of PDMS did the researchers use to culture a monolayer of cells? Did the direction of migration or other migratory behavior of the cells change depending on the shape of the PDMS stencil? According to David Kim's article, some shapes have borders that would produce different stresses upon cells so would the release of cells from these different stress conditions affect the collective migration observed?
In response to gabriel, specific cells are "labeled" as leader cells because they tend to be the first one migration. So the leader cells are just like other cells. Researchers still are unclear why these "leader" cells "lead" the migration.
In response to achung, the paper did mention that any shape can be used, but the paper used many rectangles that were parallel. So the stencil would be everything else that's not a rectangle. Migration patterns should be similar regardless of shape of the PDMS stencil. However, our project will hopefully address this issue so check back again.
All these experiments used a monolayer of epithelial cells have there been any experiments done which used a multilayered sample. Did they mention what type of epithelial cell they used? (e.g: cuboidal cells, simple columnar or simple squamous)
One question comes to mind when reading about the initial experiment: what type of medium are these cells migrating in? It's interesting to see the result of cell migration after the removal of the stencil, including the "follow the leader" migration, but I'm just curious to how closely the experimental medium resembles that of the human body. Since there are likely many more factors (such as chemical cues) in addition to merely mechanical factors that can contribute to the obvserved cell migration.
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