A mimic of the Tumor microenvironment: A simple method for generating enriched cell populations and investigating intercellular communication

Jason D. Domogauer, Sonia M. de Toledo, Edouard I. Azzam

Research output: Contribution to journalArticlepeer-review

7 Scopus citations


Understanding the early heterotypic interactions between cancer cells and the surrounding non-cancerous stroma is important in elucidating the events leading to stromal activation and establishment of the tumor microenvironment (TME). Several in vitro and in vivo models of the TME have been developed; however, in general these models do not readily permit isolation of individual cell populations, under non-perturbing conditions, for further study. To circumvent this difficulty, we have employed an in vitro TME model using a cell growth substrate consisting of a permeable microporous membrane insert that permits simple generation of highly enriched cell populations grown intimately, yet separately, on either side of the insert’s membrane for extended co-culture times. Through use of this model, we are capable of generating greatly enriched cancer-associated fibroblast (CAF) populations from normal diploid human fibroblasts following co-culture (120 hr) with highly metastatic human breast carcinoma cells, without the use of fluorescent tagging and/or cell sorting. Additionally, by modulating the pore-size of the insert, we can control for the mode of intercellular communication (e.g., gap-junction communication, secreted factors) between the two heterotypic cell populations, which permits investigation of the mechanisms underlying the development of the TME, including the role of gap-junction permeability. This model serves as a valuable tool in enhancing our understanding of the initial events leading to cancer-stroma initiation, the early evolution of the TME, and the modulating effect of the stroma on the responses of cancer cells to therapeutic agents.

Original languageEnglish (US)
Article numbere54429
JournalJournal of Visualized Experiments
Issue number115
StatePublished - Sep 20 2016

All Science Journal Classification (ASJC) codes

  • General Neuroscience
  • General Chemical Engineering
  • General Biochemistry, Genetics and Molecular Biology
  • General Immunology and Microbiology


  • Cancer biology
  • Cancer research
  • Cancer-associated fibroblasts
  • Extracellular secretion
  • Gap junctions
  • Hypoxia
  • In vitro culture model
  • Intercellular communication
  • Issue 115
  • Permeable microporous membrane insert
  • Tumor microenvironment


Dive into the research topics of 'A mimic of the Tumor microenvironment: A simple method for generating enriched cell populations and investigating intercellular communication'. Together they form a unique fingerprint.

Cite this