In the past decade, extracellular vesicles/exosomes have shown promise for the treatment of a variety of diseases and moved into the clinic with dozens of reported clinical trials globally. As the number of clinical trials increases, exosome production requires robust, scalable, and cost-effective processes. In this collaborative study with Cytiva, RoosterBio developed process parameters that can be tailored for a wide variety of therapeutic exosome programs and allows for maximum exosome recovery while maintaining quality.