Aggregate Perfusion Medium Exchange in Vertical-Wheel® Bioreactors for Scalable PSC Production
This white paper examines medium exchange strategies for scaling pluripotent stem cell (PSC) aggregate expansion in PBS Vertical-Wheel® Bioreactors. Settled medium exchange interrupts agitation and can promote aggregate merging and size heterogeneity, potentially affecting cell quality and downstream differentiation. The paper compares settled medium exchange with filter-based perfusion in PBS-0.5 Mini vessels. Perfusion maintained agitation and produced consistent growth and metabolic rates, with smaller, more spherical, and more homogeneous aggregates than settled controls. Scale-up to PBS-3 maintained comparable growth, metabolite profiles, and aggregate morphology, supporting perfusion medium exchange for scalable PSC production.
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…
How Can We Help?
Tell us what you need and we’ll take it from there.