Strategies for the Expansion of Human Induced Pluripotent Stem Cells as Aggregates in Single-Use Vertical-Wheel Bioreactors
This publication evaluates human induced pluripotent stem cell expansion as aggregates in a 0.1 L Vertical-Wheel® Bioreactor operated at a 60 mL working volume. Two commercial media, repeated-batch and fed-batch strategies, and dextran sulfate supplementation were assessed. Dextran sulfate increased maximum cell numbers across media conditions, while supplemented mTeSR1 generated the highest cell density and reduced culture duration. The optimized process achieved approximately 2.3 million cells/mL after five days with an average aggregate diameter of approximately 346 ?m. Flow cytometry, gene-expression analysis, and differentiation studies confirmed maintenance of pluripotency. The findings identify medium selection and supplementation as key process variables for efficient aggregate-based hiPSC expansion.
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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