4: Cell migration assay
Summary
Cell migration assays are used to compare mobility of cells under different conditions. It tests a cell’s ability to “crawl” across a solid substrate.
Also known as:
Scratch assay, wound-healing assay
Samples needed
A plate of confluent cells in a monolayer
Method
First, cells of interest are grown to confluence in a monolayer. Then, a pipette tip or another object is used to create a scratch through the monolayer, leaving an area of a consistent width open and free of cells. Finally, the “scratch” or “wound” is monitored and photographed to determine if one treatment group refills the gap faster than another.
Controls
The cell migration assay is nearly always used to compare mobility of cells +/- a treatment or intervention. Therefore, the usual mock treatment controls apply.
Interpretation
These microscopy images show that cells with decreased Dsg3 expression “heal the wound” faster by migrating to fill the scratched area. Therefore, the reader can conclude that one normal function of Dsg3 is to suppress cell migration. However, regardless of whether or not Dsg3 is expressed, if p38MAPK is inhibited, wound closure will not occur. Therefore, the migration-suppressing function of Dsg3 is dependent on p38MAPK activity.
Image Descriptions
Figure 1 image description:
Data from a cell migration assay.
Panel a: A grid of microscopy images of cells growing in culture. Columns show images from 0 h, 24 h, and 48 h after scratch was created. Rows from top to bottom are siNT + DMSO, siDsg3+DMSO, siNT+ SB202190, siDg3+SB202190. The top two rows both show the scratch "closing," but the siDsg3 cells close the scratch faster and more completely. With SB202190, no closure occurs.
Panel b: A line graph with time in hours on the x axis and wounding area (fold of time point 0h) on the y axis. This graph quantifies what is seen from the microscopy images in panel a. The siDsg3+DMSO condition reaches wounding area of ~0.25 by 48 hours. The siNT+DMSO condition reaches ~0.5, and both of the SB202190 conditions are still very near 1. The difference between the two DMSO conditions is significant, as is the difference between each pair of samples with DMSO vs. SB202190 with the same siRNA. ↵
Thumbnail
"Dh3-RD.jpg" ↗ by Dhifaf zeki is licensed under CC BY-SA 4.0 ↗.
Description: This is image of Scratch wound healing assay of Rhabdomyosarcoma (RD) cell line (cancer cell line) under inverted phase contrast microscope.
Author
Katherine Mattaini, Tufts University
- Rötzer, V., E. Hartlieb, J. Winkler, E. Walter, A. Schlipp, M. Sardy, V. Spindler, and J. Waschke. 2016. Desmoglein 3-dependent signaling regulates keratinocyte migration and wound healing. Journal of Investigative Dermatology 136:301-310. ↵