Optimized Serial Expansion of Human Induced Pluripotent Stem Cells Using Low-Density Inoculation to Generate Clinically Relevant Quantities in Vertical-Wheel Bioreactors
This publication describes optimization of aggregate-based human induced pluripotent stem cell expansion in single-use Vertical-Wheel® Bioreactors. Studies evaluated inoculation density, agitation, oxygen availability, nutrient supply, medium exchange, and harvest. The optimized process achieved greater than 30-fold expansion in six days with limited media use. Four serial passages produced approximately 1.06 million-fold cumulative expansion over 28 days. Cells maintained pluripotency-marker expression, normal karyotype, and the ability to generate tissues representing all three germ layers. The findings support rapid production of high-quality hiPSCs at clinically relevant quantities.
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 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…
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