A scalable 3D packaging technique for brain organoid arrays toward high-capacity bioprocessors

Abstract

Neural organoids provide a promising platform for biologically inspired computing due to their complex neural architecture and energy-efficient signal processing. However, the scalability of conventional organoid cultures is limited, restricting synaptic connectivity and functional capacity-significant barriers to developing highperformance bioprocessors. Here, we present a scalable three-dimensional (3D) packaging strategy for neural organoid arrays inspired by semiconductor 3D stacking technology. This approach vertically assembles Matrigelembedded neural organoids within a polydimethylsiloxane (PDMS)-based chamber using a removable acrylic alignment plate, creating a stable multilayer structure while preserving oxygen and nutrient diffusion. Structural analysis confirms robust inter-organoid connectivity, while electrophysiological recordings reveal significantly enhanced neural dynamics in 3D organoid arrays compared to both single organoids and two-dimensional arrays. Furthermore, prolonged culture duration promotes network maturation and increases functional complexity. This 3D stacking strategy provides a simple yet effective method for expanding the physical and functional capacity of organoid-based systems, offering a viable path toward next-generation biocomputing platforms.