Cortical-Organoid Grafted Microvascular Bed on a Chip

The self-organization of human induced pluripotent stem cells (hiPSCs) enables the development of engineered three-dimensional brain organoids, which closely mimic the intricate cytoarchitecture of the human brain. These organoids offer valuable insights into the pathogenesis of neurological diseases. However, despite their ability to replicate certain aspects of brain structure and function, they face challenges when it comes to incorporating a complex vascular network that allows for interaction with the organoids. In this study, we propose a novel approach to address this limitation by constructing a microfluidic-based, 3D cortical organoid tissue that is grafted onto a vascular bed. Our method involves three key components: (1) a perfused microvessel integrated with an extracellular matrix, allowing for dynamic flow and membrane-free culture of the endothelial layer; (2) a sprouted vascular bed network created through the use of angiogenic factors; and (3) an hiPSC-derived cortical organoid that is grafted onto the vascularized bed. Angiogenic sprouting is induced within the central graft chamber by utilizing a concentration gradient-driven angiogenic cocktail. The vascularized bed network within the microfluidic device is thoroughly characterized in terms of morphology, directional alignment under perfusion, lumen formation, and permeability. This chapter showcases the potential of our membrane-free in vitro cortical organoid grafted on a vascular bed, highlighting its utility in perfusion and scalability to a high-throughput platform.