Visualizing stromal cell dynamics in different tumor microenvironments by spinning disk confocal microscopy

Egeblad, M., Ewald, A. J., Askautrud, H. A., Truitt, M. L., Welm, B. E., Bainbridge, E., Peeters, G., Krummel, M. F., Werb, Z. (2008) Visualizing stromal cell dynamics in different tumor microenvironments by spinning disk confocal microscopy. Disease Models and Mechanisms, 1 (2-3). pp. 155-167. ISSN 17548403 (ISSN)

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DOI: 10.1242/dmm.000596


The tumor microenvironment consists of stromal cells and extracellular factors that evolve in parallel with carcinoma cells. To gain insights into the activities of stromal cell populations, we developed and applied multicolor imaging techniques to analyze the behavior of these cells within different tumor microenvironments in the same live mouse. We found that regulatory T-lymphocytes (Tregs) migrated in proximity to blood vessels. Dendriticlike cells, myeloid cells and carcinoma-associated fibroblasts all exhibited higher motility in the microenvironment at the tumor periphery than within the tumor mass. Since oxygen levels differ between tumor microenvironments, we tested if acute hypoxia could account for the differences in cell migration. Direct visualization revealed that Tregs ceased migration under acute systemic hypoxia, whereas myeloid cells continued migrating. In the same mouse and microenvironment, we experimentally subdivided the myeloid cell population and revealed that uptake of fluorescent dextran defined a low-motility subpopulation expressing markers of tumor-promoting, alternatively activated macrophages. In contrast, fluorescent anti-Gr1 antibodies marked myeloid cells patrolling inside tumor vessels and in the stroma. Our techniques allow real-time combinatorial analysis of cell populations based on spatial location, gene expression, behavior and cell surface molecules within intact tumors. The techniques are not limited to investigations in cancer, but could give new insights into cell behavior more broadly in development and disease.

Item Type: Paper
Uncontrolled Keywords: article cell hypoxia cell motion confocal microscopy human methodology neoplasm pathology stroma cell analytic method animal cell animal experiment animal model animal tissue bone marrow cell cell function cell migration cell motility cell subpopulation cell surface combinatorial chemistry controlled study dendritic cell dynamics fibroblast gene expression macrophage activation microenvironment mouse nonhuman priority journal regulatory T lymphocyte spinning disk confocal microscopy tumor cell tumor vascularization Cell Movement Humans Microscopy, Confocal Neoplasms Stromal Cells oxygen
Subjects: diseases & disorders > cancer
diseases & disorders
Investigative techniques and equipment
Investigative techniques and equipment > microscopy > flourescence microscopy
Investigative techniques and equipment > imaging
Investigative techniques and equipment > microscopy
CSHL Authors:
Communities: CSHL labs > Egeblad lab
Depositing User: Matt Covey
Date: 2008
Date Deposited: 14 Mar 2013 19:34
Last Modified: 26 Apr 2013 16:51
PMCID: PMC2562195
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