Vascularisation and Perfusion of Brain Organoids with a Meningeal Barrier on an Organ-on-Chip Platform

Brain organoids are lab-grown models of human brain development that have revolutionised neuroscience research. However, their lack of vascularisation causes hypoxia and necrosis at their core, limiting growth and maturation, and preventing the modelling of neurovascular interactions. Previous attempts to vascularise brain organoids often result in structural disruption from uncontrolled angiogenic invasion. While some strategies generate microvessels, they typically lack connection to perfusable channels, failing to support long-term tissue viability.Inspired by human embryogenesis, where vascular invasion occurs only after the developing brain is enclosed by meningeal layers, this project proposes a novel solution: enveloping the brain organoids in a meningeal barrier to protect them from aggressive vascularisation. The organoids will then be implanted in a custom-designed, perfusable organ-on-chip platform containing a pre-established vascular network of endothelial and smooth muscle cells derived from induced pluripotent stem cells (iPSCs).The work plan consists of four key steps: (1) differentiating iPSCs into neural crest-derived meningeal cells and forming a stable meningeal-brain organoid construct; (2) engineering a 3D-printed chip capable of housing and perfusing brain organoids, with a design inspired by the arteriovenous loops performed in vascular microsurgeries in vivo; (3) establishing a capillary network in the chip’s hydrogel and integrating the meningeal-brain organoid into it; and (4) perfusing the system with cell culture media to examine the effects of oxygen and nutrient delivery on organoid maturation.The anticipated outcome is a reliable, human-specific model of the neurovascular unit, capable of long-term culture and suitable for modelling cerebrovascular diseases and neurodevelopmental disorders.