High-yield Production of Extracellular Vesicle Subpopulations with Constant Quality using Batch-refeed Cultures.

The conventional manufacturing of extracellular vesicles (EVs) is characterized by low yields and batch-to-batch variability, hampering fundamental research on EVs and their practical applications. Perfusion operations have huge potential to address these limitations and increase the productivity and quality of EVs. In this study, we simulate perfusion cultures with batch-refeed systems and compare their productivity with that achieved using batch cultures. We show that a shift from batch to a batch-refeed system can increase the space-time yields of a target EV subpopulation characterized by CD81 and CD63 biomarkers by three times. Moreover, we demonstrate that the method facilitates the consistent production the target EVs from cells maintained under constant conditions for 13 days. Our results indicate that the use of perfusion cultures is a promising strategy to increase the manufacturing yield of EVs and control the production of specific EV subpopulations with constant quality attributes, thereby improving reproducibility. This article is protected by copyright. All rights reserved.