Replacement of the Flexcell Tension system

Utah State Higher Education System--University Of Utah, Salt Lake City UT

Investigators

Linked publications, trials & patents

Abstract

PROJECT SUMMARY The supplement asks for the upgrade and replacement of the current Flexcells-3000 Jr Flexcell bioreactor with a FX-6000T Flexcell system. The original application (renewal 2R01EY027920-06) proposed to test hypotheses about how TM cells navigate their response to pressure, shear, and strain, how these mechanisms are reorganized in glaucoma and how matrix stiffness and TGFb act through mechanosensitive channels to serve as triggers for fibrotic remodeling that leads to functional loss in glaucoma. Specifically, Aim 2 dissects the molecular foundation of mechanotransduction in healthy and glaucomatous TM cells with a focus on the role of extracellular matrix as a central determinant of mechanosensitivity, and tests hypotheses about involvement of TGFb signaling and mechanochannels in gene expression, intracellular stress signaling and contractility by combining electrophysiology, optical imaging, molecular analyses and stretch-evoked signaling. The legacy system currently in our lab allows the stretching of a single chamber and occasionally malfunctions, which significantly extends the experimental times, frustrates the graduate students and postdocs conducting the experiments and may impair the quality of the results. The Flexcell-6000T will improve the effectiveness and scope of present investigations by allowing us stimulate cells with uniaxial and radial strains, test 6 plates in parallel and by investigating the stretch responsiveness of cells in 3D environments.