title: Microfluidic multiculture assay to analyze biomolecular signaling in angiogenesis.
abstract: Angiogenesis (the formation of blood vessels from existing blood vessels) plays a critical role in many diseases such as cancer, benign tumors, and macular degeneration. There is a need for cell culture methods capable of dissecting the intricate regulation of angiogenesis within the microenvironment of the vasculature. We have developed a microscale cell-based assay that responds to complex pro- and antiangiogenic soluble factors with an in vitro readout for vessel formation. The power of this system over traditional techniques is that we can incorporate the whole milieu of soluble factors produced by cells in situ into one biological readout (vessel formation), even if the identity of the factors is unknown. We have currently incorporated macrophages, endothelial cells, and fibroblasts into the assay, with the potential to include additional cell types in the future. Importantly, the microfluidic platform is simple to operate and multiplex to test drugs targeting angiogenesis in a more physiologically relevant context. As a proof of concept, we tested the effect of an enzyme inhibitor (targeting matrix metalloproteinase 12) on vessel formation; the triculture microfluidic assay enabled us to capture a dose-dependent effect entirely missed in a simplified coculture assay (p < 0.0001). This result underscores the importance of cell-based assays that capture chemical cross-talk occurring between cell types. The microscale dimensions significantly reduce cell consumption compared to conventional well plate platforms, enabling the use of limited primary cells from patients in future investigations and offering the potential to screen therapeutic approaches for individual patients in vitro.
authors.affiliation: †Department of Biomedical Engineering, University of Wisconsin-Madison, Madison, Wisconsin 53705, United States.|§Department of Urology, University of Wisconsin-Madison, Madison, Wisconsin 53705, United States.|⊥Carbone Cancer Center, University of Wisconsin-Madison, Madison, Wisconsin 53705, United States.|¶Molecular and Environmental Toxicology Center, University of Wisconsin-Madison, Madison, Wisconsin 53705, United States.
authors.forename: Ashleigh B
authors.initials: AB
authors.lastname: Theberge
authors.affiliation: †Department of Biomedical Engineering, University of Wisconsin-Madison, Madison, Wisconsin 53705, United States.|⊥Carbone Cancer Center, University of Wisconsin-Madison, Madison, Wisconsin 53705, United States.
authors.forename: Jiaquan
authors.initials: J
authors.lastname: Yu
authors.affiliation: †Department of Biomedical Engineering, University of Wisconsin-Madison, Madison, Wisconsin 53705, United States.|⊥Carbone Cancer Center, University of Wisconsin-Madison, Madison, Wisconsin 53705, United States.
authors.forename: Edmond W K
authors.initials: EW
authors.lastname: Young
authors.affiliation: §Department of Urology, University of Wisconsin-Madison, Madison, Wisconsin 53705, United States.|⊥Carbone Cancer Center, University of Wisconsin-Madison, Madison, Wisconsin 53705, United States.|¶Molecular and Environmental Toxicology Center, University of Wisconsin-Madison, Madison, Wisconsin 53705, United States.
authors.forename: William A
authors.initials: WA
authors.lastname: Ricke
authors.affiliation: §Department of Urology, University of Wisconsin-Madison, Madison, Wisconsin 53705, United States.|⊥Carbone Cancer Center, University of Wisconsin-Madison, Madison, Wisconsin 53705, United States.|¶Molecular and Environmental Toxicology Center, University of Wisconsin-Madison, Madison, Wisconsin 53705, United States.
authors.forename: Wade
authors.initials: W
authors.lastname: Bushman
authors.affiliation: †Department of Biomedical Engineering, University of Wisconsin-Madison, Madison, Wisconsin 53705, United States.|⊥Carbone Cancer Center, University of Wisconsin-Madison, Madison, Wisconsin 53705, United States.|¶Molecular and Environmental Toxicology Center, University of Wisconsin-Madison, Madison, Wisconsin 53705, United States.
authors.forename: David J
authors.initials: DJ
authors.lastname: Beebe
mesh_terms: D002460:Cell Line; D060833:Cellular Microenvironment / Q000187:drug effects; D004305:Dose-Response Relationship, Drug; D042783:Endothelial Cells / Q000166:cytology / Q000187:drug effects; D004867:Equipment Design; D005347:Fibroblasts / Q000166:cytology / Q000187:drug effects; D006801:Humans; D008264:Macrophages / Q000166:cytology / Q000187:drug effects; D053508:Matrix Metalloproteinase 12 / Q000378:metabolism; D061965:Matrix Metalloproteinase Inhibitors / Q000494:pharmacology; D046210:Microfluidic Analytical Techniques / Q000295:instrumentation / Q000379:methods*; D018919:Neovascularization, Physiologic / Q000187:drug effects; D015398:Signal Transduction / Q000187:drug effects; D012995:Solubility
chemical_list: D061965:Matrix Metalloproteinase Inhibitors; D053508:Matrix Metalloproteinase 12