Additive Manufacturing of and for Organ-on-a-Chip models and analysis of their Extravesicular Secretions

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Extracellular vesicles (EVs) are tiny droplets secreted by cells and are made up of a thin lipid membrane (think of soap bubble), filled with liquid, and packed with protein and nucleic acids. EVs have been recognized to play a critical role in many diseases and function as a major communication pathway between cells and organs, acting as “parcels” sent among cells, but remain challenging to analyze. EVs, in particular those with a diameter of 60 to 120 nm, known as exosomes or small EVs, are secreted by cells, and can be retrieved from various bodily fluids, such as saliva, urine and blood. Small EVs contain specific molecular cargos, which could be used to identify their parental cells, delineate cellular composition and dysregulation, in particular for neurological diseases, cancer, and recently diabetes. EVs can migrate through the blood-brain barrier, while EVs derived from islets have been shown to contain diabetes signatures. It is already established that cancer cells secrete more EVs than healthy cells and that they can be found in blood, and contain cancer-specific signatures. Hence EVs have a high potential as accessible biomarkers for diseases and cancer. Biomanufacturing of organs-on-a-chip and miniorgans is a relatively new, rapidly evolving research topics, which enables 3D constructs of tissue, organ-on-a-chip and organoids that recapitulate relevant features of human physiology in vitro. We aim to analyze EVs secreted by complex 3D constructs and organoids, and shed light on EV signaling in health and disease. Here, we propose a platform that builds and combines the collective expertise of our team in microbioengineering and biomedical research to study EVs in the context of organoid models, including brain and hPSC-derived islet organoids. We aim to create more advanced organ models and gain understanding of EVs' role in cell-cell and organ communication during health and disease. The requested instruments include a state-of-the-art, high resolution and large build area 3D printer that allow the use of biocompatible ink for creating (i) replica molded microfluidic structures for guiding vascularization, (ii) direct bioprinting of cells to form organ-on-a-chip, and (iii) brain and islet-like organoids. The requested instruments also include a nanoparticle sizing instrument for characterizing, particularly the size distribution, EV secretions of these cellular systems.