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.
authors.affiliation: Department of Anatomy and Structural Biology, Albert Einstein College of Medicine, 1300 Morris Park Avenue, Bronx, NY, 10461, USA.
authors.forename: Ved P
authors.initials: VP
authors.lastname: Sharma
authors.forename: Brian T
authors.initials: BT
authors.lastname: Beaty
authors.forename: Dianne
authors.initials: D
authors.lastname: Cox
authors.forename: John S
authors.initials: JS
authors.lastname: Condeelis
authors.forename: Robert J
authors.initials: RJ
authors.lastname: Eddy
mesh_terms: D000818:Animals; D001681:Biological Assay / Q000379:methods*; D002450:Cell Communication / Q000187:drug effects*; D045744:Cell Line, Tumor; D002465:Cell Movement / Q000187:drug effects; D018920:Coculture Techniques; D024042:Collagen Type I / Q000737:chemistry / Q000378:metabolism; D004815:Epidermal Growth Factor / Q000494:pharmacology*; D005109:Extracellular Matrix / Q000737:chemistry / Q000378:metabolism; D005260:Female; D005353:Fibronectins / Q000737:chemistry / Q000378:metabolism; D000077156:Gefitinib; D006801:Humans; D008264:Macrophages / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D042361:Mammary Glands, Human / Q000187:drug effects* / Q000378:metabolism / Q000473:pathology; D051379:Mice; D057054:Molecular Imaging; D011799:Quinazolines / Q000494:pharmacology; D015398:Signal Transduction; D059008:Time-Lapse Imaging; D020032:Tyrphostins / Q000494:pharmacology
chemical_list: D024042:Collagen Type I; D005353:Fibronectins; D011799:Quinazolines; D020032:Tyrphostins; C101044:RTKI cpd; D004815:Epidermal Growth Factor; D000077156:Gefitinib