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    Multilayer three-dimensional filter paper constructs for the culture and analysis of aortic valvular interstitial cells

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    Author
    Sapp, Matthew C.
    Fares, Hannelle J.
    Estrada, Ana C.
    Grande-Allen, K. Jane
    Date
    2015
    Citation
    Sapp, Matthew C., Fares, Hannelle J., Estrada, Ana C., et al.. "Multilayer three-dimensional filter paper constructs for the culture and analysis of aortic valvular interstitial cells." Acta Biomaterialia, 13, (2015) 199-206. http://dx.doi.org/10.1016/j.actbio.2014.11.039.
    Published Version
    http://dx.doi.org/10.1016/j.actbio.2014.11.039
    Abstract
    Culturing aortic valvular interstitial cells in an environment that models the aortic valve is an essential step towards understanding the progression of calcific aortic valve disease. Here the adaption of a three-dimensional (3-D) stacked paper-based culture system is presented for analyzing valve cells in a thick collagen gel matrix. Filter paper layers, modeled after a 96-well plate design, were printed with a wax well-plate template and then seeded with valve cell and collagen mixtures that quickly gelled into 3-D cultures. Stacking these layers permitted extensive customization of culture thickness and cell density profiles to model the full thickness of native valve tissue. Aortic valvular interstitial cells seeded into the paper-based constructs consistently demonstrated high survival up to 14 days of culture with significant increases in cell number through the first 3 days of culture. After 4 days following seeding, valve cells in single layer cultures showed reduced smooth muscle α-actin expression with a stabilized cell density, suggesting a transition from an activated phenotype to a more quiescent state. Valve cells in multilayer cultures demonstrated the ability to migrate from layer to layer and had the highest smooth muscle α-actin expression in areas with predicted low oxygen tensions. These results establish the filter-paper-based method as a viable culture system for analyzing valve cells in an in vitro 3-D model of the aortic valve.
    Keyword
    cell activation; collagen; ECM (extracellular matrix); heart valve
    Type
    Journal article
    Citable link to this page
    http://hdl.handle.net/1911/80766
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    • Bioengineering Publications [397]
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    Managed by the Digital Scholarship Services at Fondren Library, Rice University
    Physical Address: 6100 Main Street, Houston, Texas 77005
    Mailing Address: MS-44, P.O.BOX 1892, Houston, Texas 77251-1892