Skip to main content
Biology LibreTexts

4: Cell migration assay

  • Page ID
    135769
  • \( \newcommand{\vecs}[1]{\overset { \scriptstyle \rightharpoonup} {\mathbf{#1}} } \)

    \( \newcommand{\vecd}[1]{\overset{-\!-\!\rightharpoonup}{\vphantom{a}\smash {#1}}} \)

    \( \newcommand{\id}{\mathrm{id}}\) \( \newcommand{\Span}{\mathrm{span}}\)

    ( \newcommand{\kernel}{\mathrm{null}\,}\) \( \newcommand{\range}{\mathrm{range}\,}\)

    \( \newcommand{\RealPart}{\mathrm{Re}}\) \( \newcommand{\ImaginaryPart}{\mathrm{Im}}\)

    \( \newcommand{\Argument}{\mathrm{Arg}}\) \( \newcommand{\norm}[1]{\| #1 \|}\)

    \( \newcommand{\inner}[2]{\langle #1, #2 \rangle}\)

    \( \newcommand{\Span}{\mathrm{span}}\)

    \( \newcommand{\id}{\mathrm{id}}\)

    \( \newcommand{\Span}{\mathrm{span}}\)

    \( \newcommand{\kernel}{\mathrm{null}\,}\)

    \( \newcommand{\range}{\mathrm{range}\,}\)

    \( \newcommand{\RealPart}{\mathrm{Re}}\)

    \( \newcommand{\ImaginaryPart}{\mathrm{Im}}\)

    \( \newcommand{\Argument}{\mathrm{Arg}}\)

    \( \newcommand{\norm}[1]{\| #1 \|}\)

    \( \newcommand{\inner}[2]{\langle #1, #2 \rangle}\)

    \( \newcommand{\Span}{\mathrm{span}}\) \( \newcommand{\AA}{\unicode[.8,0]{x212B}}\)

    \( \newcommand{\vectorA}[1]{\vec{#1}}      % arrow\)

    \( \newcommand{\vectorAt}[1]{\vec{\text{#1}}}      % arrow\)

    \( \newcommand{\vectorB}[1]{\overset { \scriptstyle \rightharpoonup} {\mathbf{#1}} } \)

    \( \newcommand{\vectorC}[1]{\textbf{#1}} \)

    \( \newcommand{\vectorD}[1]{\overrightarrow{#1}} \)

    \( \newcommand{\vectorDt}[1]{\overrightarrow{\text{#1}}} \)

    \( \newcommand{\vectE}[1]{\overset{-\!-\!\rightharpoonup}{\vphantom{a}\smash{\mathbf {#1}}}} \)

    \( \newcommand{\vecs}[1]{\overset { \scriptstyle \rightharpoonup} {\mathbf{#1}} } \)

    \( \newcommand{\vecd}[1]{\overset{-\!-\!\rightharpoonup}{\vphantom{a}\smash {#1}}} \)

    \(\newcommand{\avec}{\mathbf a}\) \(\newcommand{\bvec}{\mathbf b}\) \(\newcommand{\cvec}{\mathbf c}\) \(\newcommand{\dvec}{\mathbf d}\) \(\newcommand{\dtil}{\widetilde{\mathbf d}}\) \(\newcommand{\evec}{\mathbf e}\) \(\newcommand{\fvec}{\mathbf f}\) \(\newcommand{\nvec}{\mathbf n}\) \(\newcommand{\pvec}{\mathbf p}\) \(\newcommand{\qvec}{\mathbf q}\) \(\newcommand{\svec}{\mathbf s}\) \(\newcommand{\tvec}{\mathbf t}\) \(\newcommand{\uvec}{\mathbf u}\) \(\newcommand{\vvec}{\mathbf v}\) \(\newcommand{\wvec}{\mathbf w}\) \(\newcommand{\xvec}{\mathbf x}\) \(\newcommand{\yvec}{\mathbf y}\) \(\newcommand{\zvec}{\mathbf z}\) \(\newcommand{\rvec}{\mathbf r}\) \(\newcommand{\mvec}{\mathbf m}\) \(\newcommand{\zerovec}{\mathbf 0}\) \(\newcommand{\onevec}{\mathbf 1}\) \(\newcommand{\real}{\mathbb R}\) \(\newcommand{\twovec}[2]{\left[\begin{array}{r}#1 \\ #2 \end{array}\right]}\) \(\newcommand{\ctwovec}[2]{\left[\begin{array}{c}#1 \\ #2 \end{array}\right]}\) \(\newcommand{\threevec}[3]{\left[\begin{array}{r}#1 \\ #2 \\ #3 \end{array}\right]}\) \(\newcommand{\cthreevec}[3]{\left[\begin{array}{c}#1 \\ #2 \\ #3 \end{array}\right]}\) \(\newcommand{\fourvec}[4]{\left[\begin{array}{r}#1 \\ #2 \\ #3 \\ #4 \end{array}\right]}\) \(\newcommand{\cfourvec}[4]{\left[\begin{array}{c}#1 \\ #2 \\ #3 \\ #4 \end{array}\right]}\) \(\newcommand{\fivevec}[5]{\left[\begin{array}{r}#1 \\ #2 \\ #3 \\ #4 \\ #5 \\ \end{array}\right]}\) \(\newcommand{\cfivevec}[5]{\left[\begin{array}{c}#1 \\ #2 \\ #3 \\ #4 \\ #5 \\ \end{array}\right]}\) \(\newcommand{\mattwo}[4]{\left[\begin{array}{rr}#1 \amp #2 \\ #3 \amp #4 \\ \end{array}\right]}\) \(\newcommand{\laspan}[1]{\text{Span}\{#1\}}\) \(\newcommand{\bcal}{\cal B}\) \(\newcommand{\ccal}{\cal C}\) \(\newcommand{\scal}{\cal S}\) \(\newcommand{\wcal}{\cal W}\) \(\newcommand{\ecal}{\cal E}\) \(\newcommand{\coords}[2]{\left\{#1\right\}_{#2}}\) \(\newcommand{\gray}[1]{\color{gray}{#1}}\) \(\newcommand{\lgray}[1]{\color{lightgray}{#1}}\) \(\newcommand{\rank}{\operatorname{rank}}\) \(\newcommand{\row}{\text{Row}}\) \(\newcommand{\col}{\text{Col}}\) \(\renewcommand{\row}{\text{Row}}\) \(\newcommand{\nul}{\text{Nul}}\) \(\newcommand{\var}{\text{Var}}\) \(\newcommand{\corr}{\text{corr}}\) \(\newcommand{\len}[1]{\left|#1\right|}\) \(\newcommand{\bbar}{\overline{\bvec}}\) \(\newcommand{\bhat}{\widehat{\bvec}}\) \(\newcommand{\bperp}{\bvec^\perp}\) \(\newcommand{\xhat}{\widehat{\xvec}}\) \(\newcommand{\vhat}{\widehat{\vvec}}\) \(\newcommand{\uhat}{\widehat{\uvec}}\) \(\newcommand{\what}{\widehat{\wvec}}\) \(\newcommand{\Sighat}{\widehat{\Sigma}}\) \(\newcommand{\lt}{<}\) \(\newcommand{\gt}{>}\) \(\newcommand{\amp}{&}\) \(\definecolor{fillinmathshade}{gray}{0.9}\)

    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

     

    Results of a cell migration assay. Image description available.
    Figure 1. A cell migration assay in cells +/- siRNA for Dsg3 and +/- inhibitor of p38MAPK. Relevant section of caption for published figure reads: “(a) Representative bright-field images of HaCaT keratinocyte migration subjected to nontarget (siNT) or Dsg3 siRNA (siDSG3) and treated with solvent or with the p38MAPK inhibitor SB202190 (SB20). Bar is 150 mm. (b) Wound closure expressed as the remaining area uncovered by the cells. The scratch area at time point 0 hours was set to 1 (n = 4–6; *P < 0.05, #P< 0.05 vs. respective DMSO condition).” “Figure 3” by Rötzer et al.[1] [Image description]

    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


    1. 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.

    This page titled 4: Cell migration assay is shared under a CC BY-SA 4.0 license and was authored, remixed, and/or curated by Katherine Mattaini.

    • Was this article helpful?