Characterization of EGF-guided MDA-MB-231 cell chemotaxis in vitro using a physiological and highly sensitive assay system

PloS One
Verena BiswengerRoman Zantl

Abstract

Chemotactic cell migration is a central mechanism during cancer cell invasion and hence metastasis. In order to mimic in vivo conditions, we used a three-dimensional hydrogel matrix made of collagen I and a stable gradient-generating chemotaxis assay system, which is commercially available (μ-Slide Chemotaxis) to characterize epidermal growth factor (EGF)-induced chemotaxis of the human breast cancer cell line MDA-MB-231. Surprisingly, chemotactic effects of EGF on MDA-MB-231 cells could neither be observed in the standard growth medium DMEM/F-12 supplemented with 10% serum nor in starvation medium. In contrast, after adapting the cells to the serum-free growth medium UltraCULTURETM, significant chemotactic effects could be measured with high sensitivity. The extremely time-stable linear gradients, generated in the chemotaxis chamber, led to consistent directional migration of MDA-MB-231 cells. Dose-response experiments showed increased directional and kinetic response of MDA-MB-231 cells towards stable gradients of EGF. While EGF-guided directional migration (chemotaxis) was highly concentration-dependent with the highest response at 1.5 nM/mm EGF, we found that the chemokinetic effect induced by EGF was concentration-independ...Continue Reading

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Citations

Mar 27, 2020·Frontiers in Cell and Developmental Biology·Carmen WiedemannCornelia A Deeg
Aug 15, 2019·Annual Review of Cell and Developmental Biology·Michael Sheetz
Apr 9, 2020·Micromachines·Utku M SonmezLance A Davidson
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Jan 5, 2020·Bio-protocol·Sai P Visweshwaran, Alexis Gautreau
Jan 3, 2020··Alexis GautreauSai Visweshwaran

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Methods Mentioned

BETA
FCS
fluorescence correlation spectroscopy
dissect

Software Mentioned

Manager
Chemotaxis and Migration Tool
ImageJ and Migration Tool
GraphPad Prism
cellSens Dimension
GraphPad
Micro
nls
ImageJ
ImageJ Manager

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