Two-Dimensional vs. Three-Dimensional In Vitro Tumor Migration and Invasion Assays

  • Miriam Zimmermann
  • Carol Box
  • Suzanne A. Eccles
Part of the Methods in Molecular Biology book series (MIMB, volume 986)


Motility and invasion are key hallmarks that distinguish benign from malignant tumors, enabling cells to cross tissue boundaries, disseminate in blood and lymph and establish metastases at distant sites. Similar properties are also utilized by activated endothelial cells during tumor-induced angiogenesis. It is now appreciated that these processes might provide a rich source of novel molecular targets with the potential for inhibitors to restrain both metastasis and neoangiogenesis. Such therapeutic strategies require assays that can rapidly and quantitatively measure cell movement and the ability to traverse physiological barriers. The need for high-throughput, however, must be balanced by assay designs that accommodate, as far as possible, the complexity of the in vivo tumor microenvironment. This chapter aims to give an overview of some commonly used migration and invasion assays to aid in the selection of a balanced portfolio of techniques for the rapid and accurate evaluation of novel therapeutic agents.

Key words

Migration Haptotaxis Chemotaxis Invasion Motility Matrix protein 3-dimensional cultures Spheroid 







Blood–brain barrier


Basement membrane extract


Chorioallantoic membrane


Cell adhesion-mediated drug resistance


Endothelial cell


Extracellular matrix


Epidermal growth factor




Epithelial-to-mesenchymal transition


Focal adhesion kinase


Fetal calf serum




Green fluorescent protein


Hepatocellular carcinoma


Hepatocyte growth factor


Hypoxia-inducible factor-1


High-throughput (screening)


Human umbilical vein endothelial cell


Matrix metalloproteinase


Matrix metalloproteinase inhibitors




Polyethylene terephthalate


Red fluorescent protein




Squamous cell carcinoma


Tissue inhibitor of metalloproteinases


Vascular endothelial growth factor



The authors are funded by Cancer Research UK grant number C309/A8274 (S.E.) and by the Oracle Cancer Trust (M.Z. and C.B.). We acknowledge NHS funding to the NIHR Biomedical Research Centre. We thank Maria Vinci (funded by the National Centre for the Replacement, Refinement and Reduction of Animals in Research; G1000121 ID no. 94513) and other members of the Tumour Biology and Metastasis group for critical reading of the manuscript and constructive advice.


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Copyright information

© SpringerScience+Business Media New York 2013

Authors and Affiliations

  • Miriam Zimmermann
    • 1
  • Carol Box
    • 1
  • Suzanne A. Eccles
    • 1
  1. 1.Tumour Biology and Metastasis, Cancer Research UK Cancer Therapeutics Unit, Division of Cancer Therapeutics, McElwain LaboratoriesThe Institute of Cancer ResearchSurreyUK

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