Colter James, Dang Tiffany, Malinovska Julia, Corpuz Jessica May, Modrcin Dora, Krawetz Roman, Murari Kartikeya, Kallos Michael Scott
Pharmaceutical Production Research Facility (PPRF), University of Calgary, 2500 University Drive NW, Calgary, AB T2N 1N4, Canada.
Integrated Circuits and Optical Imaging Laboratory (ICOI), University of Calgary, 2500 University Drive NW, Calgary, AB T2N 1N4, Canada.
Biotechnol Rep (Amst). 2025 May 22;47:e00900. doi: 10.1016/j.btre.2025.e00900. eCollection 2025 Sep.
Human induced pluripotent stem cell (hiPSC) derived therapeutics require clinically relevant quantities of high-quality cell populations for applications in regenerative medicine. The lack of efficacy exhibited across clinical trials suggests deeper understanding of the networks governing phenotype is needed. Further, costs limit study throughput in characterizing the artificial niche relative to outcomes. We present herein an optimized strategy to enable high-throughput hiPSC expansion at <20 mL research scale. We assessed viability of single cell inoculation and aggregate preformation to facilitate proliferation. We modeled aggregate characteristics against agitation rate. Our results demonstrate tunable control with fold expansion comparable to commercial systems. Marker quantification and teratoma assay confirm functional pluripotency. This approach constitutes a scalable protocol to accelerate hiPSC research, and a significant step in advancing the rate of progress in elucidating links to derivative functionality. This work will enable statistically rigorous studies targeting hiPSC and downstream phenotype for clinical manufacturing.
人类诱导多能干细胞（hiPSC）衍生疗法在再生医学应用中需要临床上相关数量的高质量细胞群体。临床试验中表现出的疗效不足表明需要更深入地了解控制表型的网络。此外，成本限制了在表征相对于结果的人工微环境方面的研究通量。我们在此提出一种优化策略，以在小于20 mL的研究规模下实现高通量hiPSC扩增。我们评估了单细胞接种和聚集体预形成以促进增殖的可行性。我们根据搅拌速率对聚集体特征进行建模。我们的结果表明可以实现可调节的控制，其扩增倍数与商业系统相当。标志物定量和畸胎瘤试验证实了功能多能性。这种方法构成了一种可扩展的方案，以加速hiPSC研究，并在推进阐明与衍生功能联系的进展速度方面迈出了重要一步。这项工作将使针对hiPSC及其下游表型进行临床制造的统计严谨研究成为可能。
