Tumor spheroid-based migration assays for evaluation of therapeutic agents

Methods Mol Biol. 2013;986:253-66. doi: 10.1007/978-1-62703-311-4_16.


Cell migration is a key hallmark of malignant cells that contributes to the progression of cancers from a primary, localized mass to an invasive and/or metastatic phenotype. Traditional methods for the evaluation of tumor cell migration in vitro generally employ two-dimensional (2D), homogeneous cultures that do not take into account tumor heterogeneity, three-dimensional (3D) cell-cell contacts between tumor and/or host cells or interactions with extracellular matrix proteins. Here we describe a 3D tumor spheroid-based migration assay which more accurately reflects the solid tumor microenvironment and can accommodate both extracellular matrix and host cell interactions. It is a rapid and highly reproducible 96-well plate-based technique and we demonstrate its utility for the evaluation of therapeutic agents/drugs with anti-migratory properties.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Antineoplastic Agents / pharmacology*
  • Cell Adhesion
  • Cell Culture Techniques / methods
  • Cell Hypoxia
  • Cell Migration Assays / methods*
  • Cell Movement / drug effects
  • Cell Movement / physiology*
  • Cell Proliferation / drug effects
  • Cisplatin / pharmacology
  • Drug Screening Assays, Antitumor / methods*
  • Extracellular Matrix / metabolism
  • Human Umbilical Vein Endothelial Cells
  • Humans
  • Neoplasm Invasiveness
  • Neoplasms / pathology*
  • Spheroids, Cellular* / drug effects
  • Spheroids, Cellular* / metabolism
  • Spheroids, Cellular* / pathology
  • Tumor Cells, Cultured* / drug effects
  • Tumor Cells, Cultured* / metabolism
  • Tumor Cells, Cultured* / pathology
  • Tumor Microenvironment


  • Antineoplastic Agents
  • Cisplatin