Toward standardization of upscaled manufacturing of extracellular vesicles in hollow-fiber bioreactors

Abstract

Dental pulp stem cells (DPSCs) reduce acute inflammation and induce angiogenesis, primarily through paracrine factors, in which extracellular vesicles (EVs) are pivotal. However, low production yields and poor standardization have hindered clinical translation. Here, we investigated hollow-fiber bioreactor (HFB) cultivation of DPSCs to improve the scalability and standardization of DPSC-EV production. We approached this problem by investigating bioreactors with membranes of two molecular weight cut-off (MWCO) pore sizes (5 kDa and 20 kDa) and evaluating DPSC-EV production over 26 days. The quantity of EVs decreased gradually with every harvest. The 5-kDa MWCO cartridge produced a more uniform EV size distribution and marker profile compared with the 20-kDa cartridge, whereas DPSC-EVs derived from both HFBs exhibited comparable angiogenic and antiinflammatory properties. These findings demonstrate that HFB-based upscaling is a viable route for standardized production of functional DPSC-EVs upon determination of the optimal reactor parameters, thereby holding promise for advancing their clinical potential.