A microcarrier-based spheroid 3D invasion assay to monitor dynamic cell movement in extracellular matrix

Hui Liu, Tao Lu, Gert Jan Kremers, Ann L.B. Seynhaeve, Timo L.M. Ten Hagen*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

17 Citations (Scopus)


Background: Cell invasion through extracellular matrix (ECM) is a critical step in tumor metastasis. To study cell invasion in vitro, the internal microenvironment can be simulated via the application of 3D models. Results: This study presents a method for 3D invasion examination using microcarrier-based spheroids. Cell invasiveness can be evaluated by quantifying cell dispersion in matrices or tracking cell movement through time-lapse imaging. It allows measuring of cell invasion and monitoring of dynamic cell behavior in three dimensions. Here we show different invasive capacities of several cell types using this method. The content and concentration of matrices can influence cell invasion, which should be optimized before large scale experiments. We also introduce further analysis methods of this 3D invasion assay, including manual measurements and homemade semi-automatic quantification. Finally, our results indicate that the position of spheroids in a matrix has a strong impact on cell moving paths, which may be easily overlooked by researchers and may generate false invasion results. Conclusions: In all, the microcarrier-based spheroids 3D model allows exploration of adherent cell invasion in a fast and highly reproducible way, and provides informative results on dynamic cell behavior in vitro.

Original languageEnglish
Article number3
JournalBiological Procedures Online
Issue number1
Publication statusPublished - 1 Feb 2020
Externally publishedYes


  • 3D
  • Cell invasion
  • Microcarrier beads
  • Quantification
  • Spheroids
  • Time-lapse microscopy


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