title: Microsphere cytometry to interrogate microenvironment-dependent cell signaling.
abstract: Microenvironmental cues comprising surface-mediated and soluble factors control cellular signaling mechanisms underlying normal cellular responses that define homeostatic and diseased cell states. In order to measure cell signaling in single adherent cells, we developed a novel microsphere-based flow cytometry approach. Single normal or neoplastic cells were adhered to uniform microspheres that display mimetic-microenvironments comprising surface combinations of extracellular matrix (ECM) in the presence of soluble agonists/antagonists. Temporal signaling responses were measured with fluorophore-conjugated antibodies that recognize response-dependent epitopes by multiparametric flow cytometry. Using this approach we demonstrate that microenvironment-mimetic combinations of growth factors and extracellular matrix proteins generate distinct cellular signal networks that reveal unique cell signatures in normal and patient biopsy-derived neoplastic cells.
authors.affiliation: Department of Biomedicine, Center for Cancer Biomarkers, University of Bergen, Bergen, Norway. Jim.Lorens@uib.no.
authors.forename: Henriette Christie
authors.initials: HC
authors.lastname: Ertsås
authors.affiliation: Baxter Laboratory in Stem Cell Biology, Department of Microbiology and Immunology, Stanford University, Stanford, CA 94305, USA.
authors.forename: Garry P
authors.initials: GP
authors.lastname: Nolan
authors.affiliation: Life Science Division, Lawrence Berkeley National Laboratory, Berkeley, CA 94720, USA.
authors.forename: Mark A
authors.initials: MA
authors.lastname: LaBarge
authors.affiliation: Department of Biomedicine, Center for Cancer Biomarkers, University of Bergen, Bergen, Norway. Jim.Lorens@uib.no.
authors.forename: James B
authors.identifier: 0000-0002-6782-3349
authors.initials: JB
authors.lastname: Lorens
mesh_terms: D002448:Cell Adhesion; D045744:Cell Line, Tumor; D002469:Cell Separation / Q000295:instrumentation* / Q000379:methods; D004867:Equipment Design; D019544:Equipment Failure Analysis; D005109:Extracellular Matrix; D005434:Flow Cytometry / Q000295:instrumentation* / Q000379:methods; D006801:Humans; D008863:Microspheres; D009374:Neoplasms, Experimental / Q000473:pathology / Q000503:physiopathology*; D015398:Signal Transduction; D046888:Tissue Array Analysis / Q000295:instrumentation / Q000379:methods; D059016:Tumor Microenvironment / Q000502:physiology*