Abstract

Large Scale Manufacturing, Cargo Profiling, and Functional Effects of hTERT MSC EVs

Heather Branscome, Post-Doctoral Scientist, George Mason University

Ocular diseases are a major cause of visual impairment and morbidity. Furthermore, exposure to ionizing radiation (IR) can cause direct damage to the eye. Therefore, there is an urgent need to explore novel ocular therapeutics. Extracellular Vesicles (EVs) from Mesenchymal Stem Cells (MSCs) have demonstrated widespread regenerative properties across multiple pathologies. However, the reparative effects of MSC EVs against ocular damage remains relatively unexplored. Here, we report a large-scale platform for manufacturing of EVs from hTERT-immortalized MSCs and evaluate their reparative properties on retinal cells before and after exposure to IR. Additionally, we evaluate the efficacy of a novel EV lyophilization buffer for improved EV storage, transport, and stability. Physical and biochemical properties of EVs were assayed using various techniques including NTA, western blot, mass spectrometry, RNAseq, and multiplex immunoassays. EV functionality was evaluated in vitro using a combination of assays to assess cell viability, cell migration, cell cycle, and apoptosis before and after exposure to IR. Collectively, our data suggests that hTERT MSC EVs are enriched with unique cargos and that these EVs exert reparative properties on retinal cells in vitro against irradiation-induced damage. Importantly, lyophilization of EVs further extended their shelf life without impacting their function.