BACKGROUND AIMS: Autologous cell therapies using chimeric antigen receptor (CAR) T cells have shown significant clinical success in hematologic cancers. However, current production platforms face challenges in scaling up to produce sufficient numbers of cells to meet the demands of multi-dose regimens. Additionally, tight control over critical process parameters during the distinct stages of cell production is required to maximize key phenotypic characteristics of CAR T-cell products that correlate with improved clinical responses. To address these issues, we propose an integrated manufacturing process in stirred-tank bioreactors (STBs) for controlled T-cell activation and expansion. METHODS: By tailoring the stirring profile of STBs (Ambr® 15 bioreactors
Sartorius, Göttingen, Germany), microbeads functionalized with anti-CD3/CD28 antibodies allow control over the initiation/termination of T-cell activation without requiring additional washing steps to remove the activation signaling cues. RESULTS: This strategy resulted in up to a 10-fold increase in T-cell numbers compared with conventional static culture systems, resulting in a final cell concentration of 2.5 × 10 CONCLUSIONS: Overall, this approach presents a promising strategy for the scalable and tightly controlled manufacturing of T-cell therapies, particularly focusing on the T-cell activation step while minimizing manual operations, thus contributing towards more effective and cost-efficient immunotherapies.