Upscaling Human Mesenchymal Stromal Cell Production in a Novel Vertical-Wheel Bioreactor Enhances Extracellular Vesicle Secretion and Cargo Profile

This publication examines how dynamic expansion of bone marrow-derived human mesenchymal stromal cells influences extracellular vesicle production and composition. Cells were cultured on microcarriers in Vertical-Wheel bioreactors under controlled hydrodynamic conditions and compared with conventional static culture systems. Bioreactor culture increased EV secretion, enhanced expression of EV biogenesis markers, and altered EV-associated microRNA and protein cargo. Proteomic and molecular analyses identified changes in pathways related to metabolism, autophagy, oxidative stress, and cellular regulation. Scale-up studies further demonstrated successful transition between bioreactor volumes while maintaining EV productivity. The work contributes to understanding how bioprocess conditions affect MSC-derived EV manufacturing and provides valuable insight into scalable production strategies for cell-free therapeutic applications.