Comparison of technologies for manufacturing extracellular vesicles for therapeutic applications. | CU Experts

Extracellular vesicles (EVs) have gained significant attention as therapeutics, building from natural mechanisms of paracrine signaling. The field has evolved with substantial heterogeneity in methods to isolate and characterize EVs and new methods are needed to scale-up EV production for therapeutic use. In this study, we isolated EVs from four porcine donors of bone marrow-derived mesenchymal stromal cells (MSCs) via three different cell culture methods: standard tissue culture plates, a 3D printed perfusion bioreactor, and microcarriers in spinner flasks. We explored EV manufacturing yield, characteristics, and content via proteomics and RNAseq. The MSC donor and their cell culture method affected the yield of EVs produced, whereas the method of EV isolation dominated the clustering of protein and RNA contents. As a step towards therapeutic application, in vitro tubule formation and hypoxic cardiac spheroid contraction assays showed improvements in vasculogenesis and cardiac cell recovery, respectively, in the presence of EVs.

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