Analysis of Invasion Dynamics of Matrix-Embedded Cells in a Multisample Format

  • Marleen Van TroysEmail author
  • Paola Masuzzo
  • Lynn Huyck
  • Karima Bakkali
  • Davy Waterschoot
  • Lennart Martens
  • Christophe Ampe
Part of the Methods in Molecular Biology book series (MIMB, volume 1749)


In vitro tests of cancer cell invasion are the “first line” tools of preclinical researchers for screening the multitude of chemical compounds or cell perturbations that may aid in halting or treating cancer malignancy. In order to have predictive value or to contribute to designing personalized treatment regimes, these tests need to take into account the cancer cell environment and measure effects on invasion in sufficient detail. The in vitro invasion assays presented here are a trade-off between feasibility in a multisample format and mimicking the complexity of the tumor microenvironment. They allow testing multiple samples and conditions in parallel using 3D-matrix-embedded cells and deal with the heterogeneous behavior of an invading cell population in time. We describe the steps to take, the technical problems to tackle and useful software tools for the entire workflow: from the experimental setup to the quantification of the invasive capacity of the cells. The protocol is intended to guide researchers to standardize experimental set-ups and to annotate their invasion experiments in sufficient detail. In addition, it provides options for image processing and a solution for storage, visualization, quantitative analysis, and multisample comparison of acquired cell invasion data.

Key words

Cancer 3D migration Spheroid Live cell imaging Cell tracking CellMissy 



The authors acknowledge funding from the European Union’s Horizon 2020 Programme under Grant Agreement 634107 (PHC32–2014). L.H. was funded by Bijzonder Onderzoeksfonds UGent 01J04806. M.V.T. acknowledges VIB TechWatch for funding of software acquisition.


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Copyright information

© Springer Science+Business Media, LLC 2018

Authors and Affiliations

  • Marleen Van Troys
    • 1
    • 2
    Email author
  • Paola Masuzzo
    • 1
    • 2
    • 3
  • Lynn Huyck
    • 1
    • 4
  • Karima Bakkali
    • 1
  • Davy Waterschoot
    • 1
  • Lennart Martens
    • 1
    • 2
    • 3
  • Christophe Ampe
    • 1
    • 2
  1. 1.Department of Biochemistry, Faculty of Medicine and Health CareGhent UniversityGhentBelgium
  2. 2.Cancer Research Institute Ghent (CRIG)GhentBelgium
  3. 3.Medical Biotechnology Center, VIBGhentBelgium
  4. 4.UZ Gent VINRADGhentBelgium

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