A novel buoyancy technique optimizes simulated microgravity conditions for whole sensory organ culture in rotating bioreactors

Abstract

Whole-organ culture of a sensory organ in a rotating wall vessel bioreactor provides a powerful in vitro model for physiological and pathophysiological investigation as previously demonstrated for the postnatal inner ear. The model is of specific relevance as a tool for regeneration research. In the immature inner ear explant, the density was only 1.29 g/cm(3). The high density of 1.68 g/cm(3) of the functionally mature organ resulted in enhanced settling velocity and deviation from its ideal circular orbital path causing enhanced shear stress. The morphometric and physical properties, as well as the dynamic motion patterns of explants, were analyzed and numerically evaluated by an orbital path index. Application of a novel buoyancy bead technique resulted in a 6.5- to 14.8-fold reduction of the settling velocity. The deviation of the explant from its ideal circular orbital path was adjusted as indicated by an optimum value for the orbital path index (-1.0). Shear stress exerted on the inner ear explant was consequently reduced 6.4- to 15.0-fold. The culture conditions for postnatal stages were optimized, and the preconditions for transferring this in vitro model toward mature high-density stages established. This buoyancy technique may also be useful in tissue engineering of other high-density structures.