Process development and manufacture of primary human T-cells in scalable, automated stirred-tank bioreactors

Engineered Chimeric Antigen Receptor (CAR) T-cell products have recently gained FDA and EMA approval and have demonstrated significant clinical efficacy against non-Hodgkin lymphoma and pediatric B-cell acute lymphoblastic leukemia. Despite the significant clinical and commercial progress these products represent, the high costs associated with patient-specific cell therapy manufacture needs to be addressed. The work presented here focuses on the growth of human primary T-cells and CAR-T cells across a range of commercially available expansion platforms, including stirred tank bioreactors, which although routinely employed for the production of biologics, are not commonly used for the manufacture of T-cells. Initial experimental studies were carried out in an automated ambr® 250 single use bioreactor system which has demonstrated significant success for suspension and adherent mammalian cell culture applications. Building on previous work undertaken in the group which developed a new bioreactor vessel for microcarrier culture, both the new and existing bioreactor vessels were characterized with respect to cell yield, fold expansion, viability, metabolite profile, T-cell subpopulations and kLa. The comparison between the two vessels was performed based on power per unit volume, kLa and stirring speed, ranging from 100 to 200 rpm, using at least 3 different donors per condition. Please click Additional Files below to see the full abstract