Bioreactors can be used to apply fluid flow in vitro to scaffolds to improve mass
transport of media and apply mechanical forces to cells. In this study, we developed
and tested an autoclavable, modular perfusion bioreactor suitable for large scaffolds. We
investigated the effects of fluid flow induced shear stress (FFSS) on osteogenic differentiation
of human embryonic stem cell-derived mesenchymal progenitors (hES-MP
cells) cultured on large polyurethane (PU) scaffolds (30 mm diameter × 5 mm thickness)
in osteogenesis induction media (OIM). After seeding, scaffolds were either maintained
in static conditions or transferred to the bioreactor 3 days post-seeding and a continuous
flow rate of 3.47 mL/min was applied. Alkaline phosphatase activity (ALP) was used to
evaluate osteogenic differentiation and resazurin salt reduction (RR) to measure metabolic
activity after 10 days. Cultures subjected to flow contained significantly more
metabolically active cells and higher total DNA content, as well as significantly higher
ALP activity compared to scaffolds grown in static culture. These results confirm the
responsiveness of hES-MP cells to fluid flow stimuli, and present a cost-effective, userfriendly
bioreactor capable of supporting the growth and differentiation of mesenchymal
progenitor cells within scaffolds capable of filling large bone defects.

EB00000002961K

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• Design and Assessment of a Dynamic Perfusion Bioreactor for Large Bone Tissue Engineering Scaffolds