Human iPSC-derived microglia-containing 3D brain region-specific neural spheroids for disease modeling and therapeutic development 2118

Abstract Description Microglia are the primary resident immune cells of the brain. Despite accumulating evidence of the microglial involvement in neurological diseases such as Parkinson’s disease (PD), there remains a lack of physiologically relevant human neural models that recapitulate microglial contributions to physiological and pathological brain states to accelerate therapeutics discovery. To address this gap, we developed a protocol to make high throughput screening (HTS)-compatible 3D tri-culture neural spheroids via a scaffold-free aggregation of differentiated hiPSCs-derived neurons, astrocytes, and microglia. These neural spheroids mimic brain-region-specific cell-type composition and display distinct calcium oscillation that respond to compound treatment making them uniquely suitable as functional assays for drug screening. Here, we show that: (1) microglia in tri-culture spheroids exhibit a range of functional activities including phagocytosis, motility, inflammatory responses towards pathogen-associated molecules, and predictable pharmacological responses; (2) microglia incorporation alters spontaneous calcium oscillation in healthy spheroids, implicating microglia-neuron interactions; (3) healthy microglia exhibit neuroprotective functions by rescuing PD-associated functional deficit in neural spheroids. Together, we showcase a novel 3D functional tri-culture neural HTS platform for studying microglia in healthy and disease conditions which could facilitate therapeutic development. Funding Sources This research was supported in part by the Intramural/Extramural research program of the NCATS, NIH. Topic Categories Neuroimmunology (NEUR)