Metabolic Switching, Growth Kinetics and Cell Yields in the Scalable Manufacture of Stem Cell-Derived Insulin-Producing Cells
This publication examines scalable differentiation of human pluripotent stem cells into insulin-producing cells using planar CellSTACK cultures and PBS-Mini Vertical-Wheel® Bioreactors. Cells maintained greater than 80% viability and appropriate stage-specific markers throughout the seven-stage process. Early stages supported expansion and pancreatic progenitor formation, followed by reduced proliferation during endocrine differentiation. Limiting aggregate fusion improved later-stage yield. Metabolic analysis showed a shift from glycolysis toward oxidative phosphorylation, while glucose and lactate supported process characterization. The resulting cells responded to multiple secretagogues, including elevated glucose.
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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