An in vitro one-dimensional assay to study growth factor-regulated tumor cell–macrophage interaction

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5 Citations (Scopus)

Abstract

Growth factor-dependent pairing and motility between tumor cells and tumor-associated macrophages on extracellular matrix (ECM) fibers of the tumor microenvironment have been shown to enhance intravasation and metastatic spread of breast carcinomas. We describe an in vitro motility assay that combines time-lapse wide-field microscopy and micro-patterned linear adhesive substrates to reconstitute the in vivo behavior between macrophages, tumor cells, and ECM fibers in orthotopic rodent tumor models observed by intravital imaging. Commercially available linear stripes of 650 nm dye-labeled fibronectin microlithographed onto glass cover slips are sequentially plated with fluorescently labeled MTLn3 tumor cells and bone marrow-derived macrophages and time-lapse imaged for up to 8 h. Incubation with pharmacological inhibitors during the assay can identify important paracrine or autocrine signaling pathways involved in the macrophage–tumor cell interaction. This high-resolution motility assay will lead to a more detailed description of immune cell–tumor cell behavior as well as interrogating additional cell types within the tumor microenvironment which use cytokine/growth factor paracrine signaling interactions to facilitate intravasation and metastasis.

Original languageEnglish (US)
Pages (from-to)115-123
Number of pages9
JournalMethods in molecular biology (Clifton, N.J.)
Volume1172
DOIs
StatePublished - 2014
Externally publishedYes

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Intercellular Signaling Peptides and Proteins
Paracrine Communication
Tumor Microenvironment
Macrophages
Neoplasms
Extracellular Matrix
Autocrine Communication
Fibronectins
Cell Communication
Adhesives
Glass
Microscopy
Rodentia
Coloring Agents
In Vitro Techniques
Pharmacology
Breast Neoplasms
Cytokines
Neoplasm Metastasis

Keywords

  • Breast carcinoma cells
  • CSF-1
  • EGF
  • Live-cell imaging
  • Macrophage
  • Time-lapse Microscopy

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics
  • Medicine(all)

Cite this

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title = "An in vitro one-dimensional assay to study growth factor-regulated tumor cell–macrophage interaction",
abstract = "Growth factor-dependent pairing and motility between tumor cells and tumor-associated macrophages on extracellular matrix (ECM) fibers of the tumor microenvironment have been shown to enhance intravasation and metastatic spread of breast carcinomas. We describe an in vitro motility assay that combines time-lapse wide-field microscopy and micro-patterned linear adhesive substrates to reconstitute the in vivo behavior between macrophages, tumor cells, and ECM fibers in orthotopic rodent tumor models observed by intravital imaging. Commercially available linear stripes of 650 nm dye-labeled fibronectin microlithographed onto glass cover slips are sequentially plated with fluorescently labeled MTLn3 tumor cells and bone marrow-derived macrophages and time-lapse imaged for up to 8 h. Incubation with pharmacological inhibitors during the assay can identify important paracrine or autocrine signaling pathways involved in the macrophage–tumor cell interaction. This high-resolution motility assay will lead to a more detailed description of immune cell–tumor cell behavior as well as interrogating additional cell types within the tumor microenvironment which use cytokine/growth factor paracrine signaling interactions to facilitate intravasation and metastasis.",
keywords = "Breast carcinoma cells, CSF-1, EGF, Live-cell imaging, Macrophage, Time-lapse Microscopy",
author = "Sharma, {Ved P.} and Beaty, {Brian T.} and Dianne Cox and Condeelis, {John S.} and Eddy, {Robert J.}",
year = "2014",
doi = "10.1007/978-1-4939-0928-5_10",
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AU - Sharma, Ved P.

AU - Beaty, Brian T.

AU - Cox, Dianne

AU - Condeelis, John S.

AU - Eddy, Robert J.

PY - 2014

Y1 - 2014

N2 - Growth factor-dependent pairing and motility between tumor cells and tumor-associated macrophages on extracellular matrix (ECM) fibers of the tumor microenvironment have been shown to enhance intravasation and metastatic spread of breast carcinomas. We describe an in vitro motility assay that combines time-lapse wide-field microscopy and micro-patterned linear adhesive substrates to reconstitute the in vivo behavior between macrophages, tumor cells, and ECM fibers in orthotopic rodent tumor models observed by intravital imaging. Commercially available linear stripes of 650 nm dye-labeled fibronectin microlithographed onto glass cover slips are sequentially plated with fluorescently labeled MTLn3 tumor cells and bone marrow-derived macrophages and time-lapse imaged for up to 8 h. Incubation with pharmacological inhibitors during the assay can identify important paracrine or autocrine signaling pathways involved in the macrophage–tumor cell interaction. This high-resolution motility assay will lead to a more detailed description of immune cell–tumor cell behavior as well as interrogating additional cell types within the tumor microenvironment which use cytokine/growth factor paracrine signaling interactions to facilitate intravasation and metastasis.

AB - Growth factor-dependent pairing and motility between tumor cells and tumor-associated macrophages on extracellular matrix (ECM) fibers of the tumor microenvironment have been shown to enhance intravasation and metastatic spread of breast carcinomas. We describe an in vitro motility assay that combines time-lapse wide-field microscopy and micro-patterned linear adhesive substrates to reconstitute the in vivo behavior between macrophages, tumor cells, and ECM fibers in orthotopic rodent tumor models observed by intravital imaging. Commercially available linear stripes of 650 nm dye-labeled fibronectin microlithographed onto glass cover slips are sequentially plated with fluorescently labeled MTLn3 tumor cells and bone marrow-derived macrophages and time-lapse imaged for up to 8 h. Incubation with pharmacological inhibitors during the assay can identify important paracrine or autocrine signaling pathways involved in the macrophage–tumor cell interaction. This high-resolution motility assay will lead to a more detailed description of immune cell–tumor cell behavior as well as interrogating additional cell types within the tumor microenvironment which use cytokine/growth factor paracrine signaling interactions to facilitate intravasation and metastasis.

KW - Breast carcinoma cells

KW - CSF-1

KW - EGF

KW - Live-cell imaging

KW - Macrophage

KW - Time-lapse Microscopy

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