Enabling Robust and Efficient iPSC Expansion and Differentiation Processes Using High-Density Cell Banking Approaches and a Scaled-Down Computer-Controlled Parallel Vertical-Wheel Bioreactor System
This poster integrates high-density working cell banks with a computer-controlled parallel Vertical-Wheel® Bioreactor system for human induced pluripotent stem cell process development. Banks generated through three-dimensional expansion were thawed directly into suspension, reducing multiple adherent seed-train stages before differentiation. Post-thaw cultures maintained reproducible growth, OCT4 expression, normal karyotype, and functional pluripotency. The scale-down system provided dissolved oxygen and carbon dioxide control, perfusion feeding, and manufacturing-relevant hydrodynamics. Expanded cells supported definitive endoderm, cardiac, and endocrine or islet-like differentiation workflows.
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…
The PBS Closed-System Sampling Conical supports aseptic, representative sample collection from PBS Vertical-Wheel® Bioreactors while maintaining process integrity and sample quality. The single-use assembly combines a gamma-irradiated vented conical, integrated dip tube, and weldable tubing to reduce contamination risk, handling steps, and operator variability. Compatible with stem cells, immune cells, organoids, and aggregate cultures, it supports routine in-process analyses including cell counts, viability testing, metabolite monitoring, and aggregate characterization.
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