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

This publication examines how expansion of bone marrow-derived human mesenchymal stromal cells in Vertical-Wheel® Bioreactors influences extracellular vesicle (EV) production and composition. Cells were cultured on microcarriers under controlled hydrodynamic conditions and compared with 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 demonstrated consistent EV productivity across bioreactor volumes. The findings provide insight into how bioprocess conditions influence MSC-derived EV manufacturing and support development of scalable production strategies for cell-free therapeutics.