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Using the Dot Assay to Analyze Migration of Cell Sheets
Published on: December 5, 2017
Differences in adhesion and protrusion properties correlate with differences in migration speed under EGF
Yue Hou1, Sarah Hedberg, Ian C Schneider
1Department of Chemical and Biological Engineering, Iowa State University, Iowa, USA. ians@iastate.edu.
BMC Biophysics
|May 15, 2012
Summary
Epidermal growth factor (EGF) alters cell migration by changing focal adhesion (FA) properties. Fast-migrating cells exhibit lower FA intensity, suggesting specific FA dynamics are key to EGF-driven cell movement and cancer metastasis.
Area of Science:
- Cell Biology
- Biophysics
- Cancer Research
Background:
- Cell migration is crucial for biological processes like wound healing and cancer metastasis.
- Cell migration involves protrusion dynamics and focal adhesion (FA) assembly, maturation, and disassembly.
- Epidermal growth factor (EGF) is known to enhance cell migration, but its precise regulation of FA and protrusion dynamics is unclear.
Purpose of the Study:
- To investigate how FA maturation, FA dynamics, and protrusion dynamics are regulated during EGF-induced cell migration.
- To quantify FA properties and protrusion dynamics under varying doses of EGF stimulation using advanced microscopy techniques.
Main Methods:
- Utilized total internal reflection fluorescence (TIRF) microscopy to observe cell behavior.
- Employed image analysis to quantify focal adhesion (FA) properties and protrusion dynamics.
- Stimulated cells with different concentrations of Epidermal Growth Factor (EGF).
Main Results:
- EGF broadened cell migration rate distribution, creating faster and slower cells.
- FA intensity decreased with increasing EGF concentration; FA number peaked at intermediate concentrations.
- Fast-migrating cells showed lower FA intensity, and protrusion waves correlated with faster migration.
Conclusions:
- FA properties and protrusion dynamics correlating with migration speed do not always correlate with EGF stimulation.
- Uncorrelated FA dynamics are sensitive indicators of differential cellular responses to EGF.
- EGF's ability to modulate individual cell migration speeds has implications for cancer metastasis, potentially selecting for more invasive subpopulations.
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