Engineering neuronal networks in granular microgels to innervate bioprinted cancer organoids on-a-chip

Organoid models are invaluable for studying organ processes in vitro, offering an unprecedented ability to replicate organ function. Despite recent advancements that have increased their cellular complexity, organoids generally lack key specialized cell types, such as neurons, limiting their ability to fully model organ function and dysfunction. Innervating organoids remains a significant challenge due to the asynchronous biological cues governing neural and organ development. Here, we present a versatile organ-on-a-chip platform designed to innervate organoids across diverse tissue types. Our strategy enables the development of innervated granular hydrogel tissue constructs, followed by the sequential addition of organoids. The microfluidic device features an open tissue chamber, which can be easily manipulated using standard pipetting or advanced bioprinting techniques. Engineered to accommodate microgels of any material larger than 50 μm, the chamber provides flexibility for constructing customizable hydrogel environments. Organoids and other particles can be precisely introduced into the device at any stage using aspiration-assisted bioprinting. To validate this platform, we demonstrate the successful growth of primary mouse superior cervical ganglia (mSCG) neurons and the platform's effectiveness in innervating prostate cancer spheroids and patient-derived renal cell carcinoma organoids. This platform offers a robust and adaptable tool for generating complex innervated organoids, paving the way for more accurate in vitro models of organ development, function, and disease.