Overcoming Bioprocess Bottlenecks Through Advanced Bioreactor Process Development

This publication investigates strategies to overcome major manufacturing bottlenecks associated with large-scale expansion of human induced pluripotent stem cells (hiPSCs) using Vertical-Wheel bioreactors. Computational fluid dynamics modeling was used to characterize hydrodynamic conditions and guide development of scalable inoculation, expansion, harvesting, and serial-passaging protocols. The study demonstrated successful single-cell inoculation, achieving greater than 30-fold expansion within six days under optimized fed-batch conditions while maintaining homogeneous aggregate morphology. An in-vessel aggregate dissociation protocol achieved more than 95% harvest efficiency and enabled serial passaging and scale-up into larger bioreactor volumes. Expanded cells maintained normal karyotype, expression of pluripotency markers, and functional differentiation into cell types representing all three germ layers. The work provides a framework for scalable clinical and manufacturing production of high-quality hiPSCs.