Scaled preparation of extracellular vesicles from conditioned media

Extracellular vesicles (EVs) especially of mesenchymal stem/stomal cells (MSCs) are increasingly considered as biotherapeutic agents for a variety of different diseases. For translating them effectively into the clinics, scalable production processes fulfilling good manufacturing practice (GMP) are needed. Like for other biotherapeutic agents, the manufacturing of EV products can be subdivided in the upstream and downstream processing and the subsequent quality control, each of them containing several unit operations. During upstream processing (USP), cells are isolated, stored (cell banking) and expanded; furthermore, EV-containing conditioned media are produced. During downstream processing (DSP), conditioned media (CM) are processed to obtain concentrated and purified EV products. CM are either stored until DSP or are directly processed. As first unit operation in DSP, clarification removes remaining cells, debris and other larger impurities. The key operations of each EV DSP is volume-reduction combined with purification of the concentrated EVs. Most of the EV preparation methods used in conventional research labs including differential centrifugation procedures are limited in their scalability. Consequently, it is a major challenge in the therapeutic EV field to identify appropriate EV concentration and purification methods allowing scale up. As EVs share several features with enveloped viruses, that are used for more than two decades in the clinics now, several principles can be adopted to EV manufacturing. Here, we introduce and discuss volume reducing and purification methods frequently used for viruses and analyze their value for the manufacturing of EV-based therapeutics. (c) 2021 Elsevier B.V. All rights reserved.

Automated summary

Extracellular vesicles (EVs), especially those from mesenchymal stem/stromal cells (MSCs), are being increasingly explored as biotherapeutic agents for various diseases. To translate them effectively into clinical applications, scalable production processes that comply with good manufacturing practice (GMP) are necessary. The scalability of conventional EV preparation methods poses a major challenge in the therapeutic EV field, requiring the identification of appropriate concentration and purification methods for scale-up. Adopting principles from enveloped viruses, which have been used in clinics for over two decades, can provide valuable insights for EV manufacturing.