Comparative Assessment of Whole Organ Tissue Processing Methods for the Isolation of Extracellular Vesicles From...

Extracellular vesicles (EVs) are small anuclear cellular membrane encapsulated fragments of importance for cellular interaction and transfer of information. These small vesicles, diverse in size and functionality, can be obtained from cells, tissues and bodily fluids. A complicated step for obtaining EVs from whole organs is understanding the optimal methodology for organ processing. In this study, we have examined two different techniques: one enzymatic and one novel non-enzymatic automated tissue dissociation (ATD) machine. Animals were perfused, organs extracted, and techniques comparatively applied. We have used these techniques for organ-based dissociation followed by EV isolation from the dissociated tissues (heart, kidney, lung). While both approaches allow isolation of intact EVs there are distinct differences in overall cell and particle yields. Our study highlights tissue specific inter-organ variability and differential impact of dissociation strategies on organ-based EV profiles, as well as cellular characteristics. Our findings indicate that EV yields and characteristics varies between enzymatic and ATD techniques as well as between organs with highest EV yield obtained from kidneys following enzymatic dissociation. Our findings can be rapidly transferred to other setups or developed to enable enumeration and characterization of EVs obtained from whole organs in physiological and pathological settings.