Fabrication of Cartilage Sheets

Cartilage sheets were fabricated as previously described (9 (link)), with minor modifications. Briefly, chondrocytes from second passage were typsinized and plated at 1.9 × 10⁶ cells/cm² into custom stainless steel biochambers consisting of a 4.0 × 4.0 cm upper chamber with a porous (10 μm) base consisting of a porous polyester membrane (PET1009030; Sterlitech, Kent, WA) coated with human fibronectin (BD Biosciences, San Jose, CA), which suspended the cells above the lower chamber which was either a 100 mm tissue culture plate or a custom cylindrical chamber with sufficient capacity to allow the addition of sufficient medium volume to totally immerse the biochamber (~300 ml). The large container allowed the cells in the upper chamber to be exposed to much more medium, thus avoiding medium depletion, as evinced by yellowing medium.