Using Microarrays to Interrogate Microenvironmental Impact on Cellular Phenotypes in Cancer

J Vis Exp. 2019 May 21:(147):10.3791/58957. doi: 10.3791/58957.

Abstract

Understanding the impact of the microenvironment on the phenotype of cells is a difficult problem due to the complex mixture of both soluble growth factors and matrix-associated proteins in the microenvironment in vivo. Furthermore, readily available reagents for the modeling of microenvironments in vitro typically utilize complex mixtures of proteins that are incompletely defined and suffer from batch to batch variability. The microenvironment microarray (MEMA) platform allows for the assessment of thousands of simple combinations of microenvironment proteins for their impact on cellular phenotypes in a single assay. The MEMAs are prepared in well plates, which allows the addition of individual ligands to separate wells containing arrayed extracellular matrix (ECM) proteins. The combination of the soluble ligand with each printed ECM forms a unique combination. A typical MEMA assay contains greater than 2,500 unique combinatorial microenvironments that cells are exposed to in a single assay. As a test case, the breast cancer cell line MCF7 was plated on the MEMA platform. Analysis of this assay identified factors that both enhance and inhibit the growth and proliferation of these cells. The MEMA platform is highly flexible and can be extended for use with other biological questions beyond cancer research.

Publication types

  • Research Support, N.I.H., Extramural
  • Video-Audio Media

MeSH terms

  • Extracellular Matrix / metabolism
  • Extracellular Matrix Proteins / metabolism
  • Humans
  • Ligands
  • MCF-7 Cells
  • Microarray Analysis / methods*
  • Neoplasms / metabolism
  • Neoplasms / pathology*
  • Phenotype
  • Tumor Microenvironment*

Substances

  • Extracellular Matrix Proteins
  • Ligands