Senior Research Scientist Corning Life Sciences, Taiwan (Republic of China)
Abstract: The therapeutic benefits of mesenchymal stem cells (MSCs) are widely believed to be derived from their paracrine signaling, such as extracellular vesicles (EVs). Cell-free therapy using EVs is an active and emerging field in regenerative medicine. Despite the great potential of MSC-derived small extracellular vesicles (sEVs), a major challenge for EV production in manufacturing is the choice of culture system. Among the biophysical strategies, 3D and dynamic cultures are proposed to enhance their therapeutic efficacy, although the mechanisms are not yet fully understood. In this study, we aimed to better understand the effects and mechanisms of these culture microenvironments on MSC-derived EVs during EV production. The experimental results showed that dynamic culture induced higher EVs particles secretion from NTA data, while 3D spheroid culture, in contrast, mitigated EV quantity. Furthermore, more informative biomolecules such as proteins and miRNAs were encapsulated per EV particle with 3D spheroid culture, in both dynamic and static cultures. Our hypothesis is that shear force triggers more EV particles, albeit with less surface marker expression, and the 3D culture environment stimulates more surface marker proteins to be packed on each EV. In summary, the results suggest that MSCs secrete more informative EVs from 3D cultures, and dynamic cultures trigger more EV production but compromise the quality of EVs. This study provides new insights into MSC-EVs obtained from different culture environments, which should be considered and justified in EV manufacturing.
