Scaling-Up Vertical Wheel Bioreactors Based on Cell Aggregate Exposure to Shear Stress and Energy Dissipation Rate
This publication investigates hydrodynamic scale-up of human pluripotent stem cell aggregate cultures in 100 mL and 500 mL Vertical-Wheel® Bioreactors. Three-dimensional flow simulations and aggregate trajectory analysis were used to evaluate shear stress and energy dissipation rate exposure across multiple agitation conditions. Exposure profiles were compared with aggregation efficiency, cell growth, aggregate size, and fold expansion during six-day cultures. Energy dissipation rate exposure correlated with aggregate formation and expansion performance, while shear stress was not a significant predictor under the tested conditions. The study proposes trajectory-based energy dissipation exposure as a more representative scale-up parameter than maximum or volume-averaged metrics for aggregate-based stem cell manufacturing.
This publication reports a fully suspended process for expansion and six-stage differentiation of human induced pluripotent stem cells into stem cell-derived islets in 0.1 L and 0.5 L Vertical-Wheel® Bioreactors. A fivefold increase in vessel volume produced a twelvefold increase in islet-equivalent yield without compromising islet structure. The 27-day process generated endocrine populations expressing ?-, ?-, and ?-cell markers with glucose-responsive insulin secretion and transcriptomic characteristics approaching adult islets. The study also evaluated donor-derived iPSC lines, cellular…
This protocol provides step-by-step guidance for forming and expanding pluripotent stem cell aggregates in PBS-MINI Vertical-Wheel® bioreactors. The workflow begins with thawing and adherent seed-train culture in T-flasks, followed by transfer to three-dimensional suspension culture in 0.1 L or 0.5 L vessels. Instructions cover inoculation, medium exchange, sampling, morphology assessment, metabolite and pH analysis, cell counting, viability testing, and harvest in the PBS-Mini bioreactor. The document presents recommended starting parameters for seeding density, working volume, agitation…
This protocol provides detailed instructions for expanding mesenchymal stem cells on microcarriers using PBS-Mini bioreactors with Vertical-Wheel technology. The document covers the complete workflow from recovery of cryopreserved MSCs and expansion in adherent culture through inoculation, suspension culture, sampling, medium exchange, and harvest in three-dimensional bioreactor systems the PBS-Mini bioreactor. Recommended process parameters are based on internal optimization studies using multiple MSC lines and media formulations. Guidance is provided for microcarrier preparation, cell…
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