Moving upwards: A simple and flexible in vitro three-dimensional invasion assay protocol

Tanner J. McArdle, Brenda M. Ogle, Felicite K. Noubissi

Research output: Contribution to journalArticlepeer-review

1 Scopus citations


Although 3D invasion assays have been developed, the challenge remains to study cells without affecting the integrity of their microenvironment. Traditional 3D assays such as the Boyden Chamber require that cells are displaced from the original culture location and moved to a new environment. Not only does this disrupt the cellular processes that are intrinsic to the microenvironment, but it often results in a loss of cells. These problems are especially challenging when dealing with cells that are either rare, or extremely sensitive to their microenvironment. Here, we describe the development of a 3D invasion assay that avoids both concerns. In this assay, cells are plated within a small well and an ECM matrix containing a chemoattractant is laid atop the cells. This requires no cell displacement, and allows the cells to invade upwards into the matrix. In this assay, cell invasion as well as cell morphology can be assessed within the collagen gel. Using this assay, we characterize the invasive capacity of rare and sensitive cells; the hybrid cells resulting from fusion between breast cancer cells MCF7 and mesenchymal/ multipotent stem/stroma cells (MSCs).

Original languageEnglish (US)
Article numbere56568
JournalJournal of Visualized Experiments
Issue number133
StatePublished - Mar 12 2018

Bibliographical note

Funding Information:
This work was supported by the National Institutes of Health (NIMHD-G12MD007581) through the RCMI-Center for Environmental Health at Jackson State University and the U.S. Department of Defense, Idea Award 11-1-0205.

Publisher Copyright:
© 2018 Journal of Visualized Experiments.


  • Cancer research
  • Invasion
  • Inverted
  • Issue 133
  • Microenvironment
  • Migration
  • Rare
  • Sensitive
  • Three-dimensional (3D)


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