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

摘要: Culturing aortic valvular interstitial cells in an environment that models the valve is essential step towards understanding progression of calcific disease. Here adaption a three-dimensional (3-D) stacked paper-based culture system presented for analyzing thick collagen gel matrix. Filter paper layers, modeled after 96-well plate design, were printed with wax well-plate template and then seeded cell mixtures quickly gelled into 3-D cultures. Stacking these layers permitted extensive customization thickness density profiles to model full native tissue. Aortic constructs consistently demonstrated high survival up 14 days significant increases number through first 3 culture. After 4 following seeding, single layer cultures showed reduced smooth muscle α-actin expression stabilized density, suggesting transition from activated phenotype more quiescent state. Valve multilayer ability migrate had highest areas predicted low oxygen tensions. These results establish filter-paper-based method as viable vitro valve.