Fig. (1) The rotating wall vessel (RWV). Panel A: The rotating wall vessel serves as a low fluid shear cell culture apparatus. The vessel is completely filled with media containing cells (no air bubbles) and rotated at 25 rpm. In the pictured orientation of the vessel (LSMMG condition), this creates a low shear, low turbulence suspension culture environment for cell growth. Aeration is provided via an air-permeable membrane located in the back of the vessel (not visible). Panel B: The two orientations of the RWV for experimental set up. For RWV experiments, two separate apparatuses are used that are set up in two different orientations using the same culture inoculation. In the control orientation, the low fluid shear environment is abolished and increased fluid shear levels are present. The only difference in the two cultures is the orientation of each vessel. Panel C: A diagram of the orbital path of a representative cell in the LSMMG orientation of the RWV to demonstrate the nature of the low fluid shear cell suspension. This is achieved in large part by the solid body rotation of the media in the apparatus in such a way that sedimentation is offset to allow suspension with a minimum of fluid shear forces.