(183n) Bioprocess Sensor for iPSC Culture Online Monitoring and Automation

Regenerative medicine is projected to reach a global market of nearly $200 billion, yet the production of live cell-based therapies, particularly those involving induced pluripotent stem cells (iPSCs), still faces major challenges. Scalable and reliable iPSC manufacturing strategies are urgently needed, as their success is closely tied to the broader clinical application of regenerative therapies. However, iPSC productivity and functional identity are highly sensitive to culture conditions. Culture dynamics significantly influence critical process outcomes, including cost, growth rate, productivity—measured as integrated viable cell density (IVCD)—and, most importantly, pluripotency, which serves as a key critical quality attribute (CQA). Even subtle variations in culture environments can trigger cell stress, leading to heterogeneity and a decline in pluripotency. Given the complexity of iPSCs, it is essential to gain a comprehensive understanding of their metabolism, closely monitor their metabolic state, and guide them along the desired expansion trajectory. In this study, we employed a two-photon excitation (TPE) sensor for real-time, non-invasive monitoring of redox ratio changes. This approach enabled the acquisition of operational data, and the identification of amino acids correlated with cell reproducibility, informing more precise feeding strategies. Specifically, tryptophan, alanine, glutamine, and phenylalanine were identified as key contributors to media optimization. By balancing the consumption of essential amino acids and the synthesis of non-essential ones, we achieved improved cell quality, offering a promising direction for scalable and efficient iPSC expansion.