A cancer cell spheroid assay to assess invasion in a 3D setting

Eric B. Berens, Jon M. Holy, Anna T. Riegel, Anton Wellstein

Research output: Contribution to journalArticlepeer-review

91 Scopus citations


The invasive nature of cancer cell lines is thought to correlate with their metastatic potential. Most traditional assays, however, do not examine these invasive features in a three-dimensional environment and the resulting data suffer from reduced biological applicability. Here an approach is presented to visualize the invasive ability of cell lines in a physiologically relevant setting. The cancer cell spheroid invasion assay first utilizes gravity to generate spheroids within drops of media that hang from the lid of a cell culture dish. Next, these spheroids are embedded in a 3D matrix consisting of a mixture of basement membrane materials and type I collagen. Cancer cell egression from the spheroids into the surrounding matrix is then monitored over time. The method described here can be modified to examine invasion after coculture of different cell types, inclusion of drugs/inhibitors, or alterations in extracellular matrix (ECM) constituents.

Original languageEnglish (US)
Article numbere53409
JournalJournal of Visualized Experiments
Issue number105
StatePublished - Nov 20 2015

Bibliographical note

Publisher Copyright:
© 2015 Journal of Visualized Experiments.


  • 3D
  • Basement membrane
  • Cancer
  • Collagen
  • Hanging drop
  • Invasion
  • Issue 105
  • Matrigel
  • Medicine
  • Metastasis
  • Sphere
  • Spheroid


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