The ability to quantify cell migration and invasion is critical in the study of cancer metastasis. Current invasion assays, such as the Boyden Chamber, present difficulties in the measurement of the invasion of cells that are few in number and are intrinsically tied to the cell microenvironment. There exists a need for a three-dimensional invasion assay that is easily reproduced, accessible for most laboratories, and requires no displacement of cells from their original microenvironment. Here we present a simple design for an inverted vertical invasion assay able to assess the invasion capabilities of cells in a three dimensional, extracellular matrix-based environment without displacement from the original culture location. We used the assay to determine the migratory capacity of hybrids between mesenchymal/multipotent stem/stroma cells (MSCs) and breast cancer cells MCF7. These hybrids are formed reliably but rarely (1 in 1,000 cells) and for this reason require an invasion assay that does not involve extensive cell manipulation. Using this assay, we found that MSCs, breast cancer cells, and corresponding fusion products are able to migrate and invade through the extracellular matrix and that hybrids invade in a manner more similar to stromal cells than cancer cells. Thus, this assay can aid the study of the invasive capacity of both cancerous cells and associated fusion hybrids and could augment testing of therapeutic strategies to inhibit metastatic spread.
Bibliographical noteFunding Information:
The authors thank Dr. Peiman Hematti (Division of Hematology and Oncology, University of Wisconsin, Madison, WI, USA) for the gift of MSCs, Dr. Thomas Kerppola (Professor of Biological Chemistry, University of Michigan, Ann Arbor, MI, USA) for the gift of BiFC constructs, Dr. Philip Jung (University of Minnesota, Minneapolis, MN, USA) for help with ANOVA with Tukey?s HSD post hoc test, the MPSCM, and the 3D image reconstruction. MPLSM was performed in the lab of Dr. Paulo Provenzano and the confocal microscope imaging in the lab of Dr. Patrick Alford (University of Minnesota, Minneapolis, MN, USA). This work was supported by U.S. Department of Defense, Idea Award 11-1-0205, the University of Minnesota Undergraduate Research Opportunities Program, and the National Institutes of Health (NIMHD-G12MD007581) through the RCMI-Center for Environmental Health at Jackson State University.
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- 3D assay