• A parallelized 8-chamber rotary-perfusion bioreactor was successfully developed. • CFD identified 0.03 m/s and 3-6 rpm as optimal for low-shear dECM suspension. • The system maintains stable scaffold suspension with shear stress below 0.2 Pa. • Dynamic culture significantly improved MSC viability and 3D network organization. • The platform enables high-throughput, low-cost fabrication of complex organoids. The scalable and physiological fabrication of organoids remains a pivotal challenge in tissue engineering. Decellularized extracellular matrix (dECM) offers an ideal, organ-specific biomaterial scaffold, yet its full potential is often unrealized in static culture due to limited mass transport and poorly controlled mechanical environments. Here, we present a parallelized rotary-perfusion bioreactor designed to address these limitations by uniquely integrating simulated microgravity with low-shear perfusion-a combination that enables true three-dimensional suspension while avoiding the detrimental hydrodynamic forces associated with conventional dynamic systems. The system integrates eight independent culture chambers with precise control over perfusion and rotation, allowing for high-throughput, low-volume, and co-culture-compatible organoid production. Using computational fluid dynamics (CFD) coupled with experimental validation, we systematically optimized the operating parameters to achieve a stable, low-shear microenvironment ( τ e q , m a x < 0.3 P a , perfusion velocity 0.03 m/s, rotation speed 3-6 rpm). Biological validation using human umbilical cord mesenchymal stem cells cultured on murine dECM demonstrated that, compared to static controls, dynamic culture significantly enhanced cell viability (10 % higher on average), proliferation, and cytoskeletal organization over five days. This work establishes a robust, scalable platform for low-loading dynamic culture of cell-seeded dECM constructs, providing a foundation for subsequent organoid maturation under controlled rotation-perfusion conditions. By decoupling shear control from perfusion and integrating simulated microgravity, our approach addresses a key technological gap and offers a generally applicable strategy for engineering physiologically relevant 3D tissue models. The system holds promising potential for advancing tissue engineering, drug screening, and regenerative medicine applications.
Sun et al. (Wed,) studied this question.