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An In Vitro One-Dimensional Assay to Study Growth Factor-Regulated Tumor Cell–Macrophage Interaction

  • Ved P. Sharma
  • Brian T. Beaty
  • Dianne Cox
  • John S. Condeelis
  • Robert J. Eddy
Part of the Methods in Molecular Biology book series (MIMB, volume 1172)

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.

Key words

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

Notes

Acknowledgment

We thank the people from the Condeelis, Segall, and Cox laboratories for helpful discussions. This work was supported by CA150344 (R.J.E. and V.P.S.), CA100324 (J.S.C. and D.C.) and a postdoctoral fellowship from Susan G. Komen for the Cure© KG111405 to V.P.S.

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Copyright information

© Springer Science+Business Media, New York 2014

Authors and Affiliations

  • Ved P. Sharma
    • 1
    • 2
  • Brian T. Beaty
    • 1
  • Dianne Cox
    • 1
    • 2
  • John S. Condeelis
    • 1
    • 2
  • Robert J. Eddy
    • 1
  1. 1.Department of Anatomy and Structural BiologyAlbert Einstein College of MedicineBronxUSA
  2. 2.Gruss Lipper Biophotonics CenterAlbert Einstein College of MedicineBronxUSA

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