title: Novel multi-functional fluid flow device for studying cellular mechanotransduction.
abstract: Cells respond to their mechanical environment by initiating multiple mechanotransduction signaling pathways. Defects in mechanotransduction have been implicated in a number of pathologies; thus, there is need for convenient and efficient methods for studying the mechanisms underlying these processes. A widely used and accepted technique for mechanically stimulating cells in culture is the introduction of fluid flow on cell monolayers. Here, we describe a novel, multifunctional fluid flow device for exposing cells to fluid flow in culture. This device integrates with common lab equipment including routine cell culture plates and peristaltic pumps. Further, it allows the fluid flow treated cells to be examined with outcomes at the cell and molecular level. We validated the device using the biologic response of cultured UMR-106 osteoblast-like cells in comparison to a commercially available system of laminar sheer stress to track live cell calcium influx in response to fluid flow. In addition, we demonstrate the fluid flow-dependent activation of phospho-ERK in these cells, consistent with the findings in other fluid flow devices. This device provides a low cost, multi-functional alternative to currently available systems, while still providing the ability to generate physiologically relevant conditions for studying processes involved in mechanotransduction in vitro.
authors.affiliation: Department of Orthopaedics, University of Maryland School of Medicine, Baltimore, MD 21201, USA.
authors.forename: James S
authors.initials: JS
authors.lastname: Lyons
authors.affiliation: Department of Orthopaedics, University of Maryland School of Medicine, Baltimore, MD 21201, USA.
authors.forename: Shama R
authors.initials: SR
authors.lastname: Iyer
authors.affiliation: Department of Orthopaedics, University of Maryland School of Medicine, Baltimore, MD 21201, USA.
authors.forename: Richard M
authors.initials: RM
authors.lastname: Lovering
authors.affiliation: University of Maryland School of Nursing, Baltimore, MD 21201, USA.
authors.forename: Christopher W
authors.initials: CW
authors.lastname: Ward
authors.affiliation: Department of Orthopaedics, University of Maryland School of Medicine, Baltimore, MD 21201, USA. Electronic address: jstains@som.umaryland.edu.
authors.forename: Joseph P
authors.initials: JP
authors.lastname: Stains
mesh_terms: D000818:Animals; D020013:Calcium Signaling; D018929:Cell Culture Techniques / Q000295:instrumentation; D002478:Cells, Cultured; D003592:Cytophotometry / Q000295:instrumentation*; D057446:Hydrodynamics; D040542:Mechanotransduction, Cellular; D010006:Osteoblasts / Q000502:physiology; D051381:Rats; D059010:Single-Cell Analysis / Q000295:instrumentation
keywords: Bone cells; Fluid shear stress; Intracellular calcium; Mechanotransduction; Osteoblast; Osteocyte; Signal transduction