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Fluid Mechanics, Cell Distribution, and Environment in Cell Cube Bioreactors

Cultivation of MRC‐5 cells and attenuated hepatitis A virus (HAV) for the production of VAQTA, an inactivated HAV vaccine ( 1 ), is performed in the Cell Cube reactor, a laminar flow fixed‐bed bioreactor with an unusual diamond‐shaped, diverging‐converging flow geometry. These disposable bioreactors have found some popularity for the production of cells and gene therapy vectors at intermediate scales of operation ( 2 , 3 ). Early testing of the Cell Cube revealed that the fluid mechanical environment played a significant role in nonuniform cell distribution patterns generated during the cell growth phase. Specifically, the reactor geometry and manufacturing artifacts, in combination with certain inoculum practices and circulation flow rates, can create cell growth behavior that is not simply explained. Via experimentation and computational fluid dynamics simulations we can account for practically all of the observed cell growth behavior, which appears to be due to a complex mixture of flow distribution, particle deposition under gravity, fluid shear, and possibly nutritional microenvironment.