Scalable Differentiation of Hematopoietic Progenitor Cells from hiPSCs Using a Bioreactor System

A method for scalable and efficient differentiation of hematopoietic progenitor cells (HPCs) from human induced pluripotent stem cells (hiPSCs) represents a pivotal advancement for cell-based therapies targeting cancer, autoimmune disorders, and hematological diseases. Although effective for small-scale production, traditional static culture methods are limited in scalability and reproducibility, necessitating the development of bioreactor-based approaches. In this study, we optimized HPC differentiation from hiPSC lines using a 30-mL stirred suspension bioreactor. The obtained embryoid bodies expanded from ~ 100 µm to 400 µm between days 3 and 13, while their number decreased through fusion. OCT4 expression declined from day 3, followed by transient BRACHYURY and MIXL1 expression, and sequential induction of CD34 from day 7 and CD43 from day 10. Flow cytometry analysis revealed that CD34⁺ cells peaked at 63.3% on days 9–10, while CD34⁺CD43⁺ cells increased markedly by day 13. The total cell count reached 1.2×10⁸ by day 10, yielding 9.7×10⁷ CD34⁺ cells. MM11 hiPSCs differentiated more efficiently than 201B7 hiPSCs, indicating cell line-specific differences. Compared with static culture, this bioreactor system achieved superior yield, uniformity, and scalability. These findings validate the utility of stirred bioreactors for clinical-scale HPC production and provide a foundation for process optimization and translational development.