Multiscale 3D microfluidic platform for intraorganoid delivery

Neural organoids are emerging as advanced three-dimensional (3D) in vitro models for recapitulating human development and pathology, but they lack dedicated mass-transport pathways that perfuse interior regions, limiting control over solute concentrations in deep tissues. Microfluidic technologies hold promise for intra-organoid delivery, yet creating high-resolution 3D transport architectures spanning arteriole-to-venule scales while integrating them with minimal disruption to morphogenesis remains challenging. Here, we introduce a multiscale 3D microfluidic delivery platform that embeds lithographically defined, flexible, thread-like microchannels into organoids during growth. The nanoporous interface along the embedded microchannels enables controlled diffusive transport of biomolecules into localized regions with ~100 μm spatial resolution and to depths of ~400 μm from the organoid center, with minute-scale temporal precision, as demonstrated with dyes, morphogens, and MRI contrast agents. Delivery of growth factor-supplemented media leads to reduced apoptosis near the microchannels and improved neural tissue integrity. This platform offers a robust means to interrogate deep, site-specific microenvironments and to advance studies of organoid viability, structural organization, and functional maturation.