Protocol for studying dynamics of parasite-host-mediated endothelial inflammation in 3D engineered brain microvessels

Bioengineered microvessels offer a controllable human system over cellular composition, vessel architecture, flow, and circulating components for studying vascular diseases. This protocol details the use of 3D brain microvessels to model in vivo-like parasite binding, maturation, and induced vascular inflammation during cerebral malaria progression. We outline steps for studying microvascular inflammation through immunofluorescence, ultrastructural imaging, RNA sequencing, and functional assays for permeability and leukocyte recruitment. This protocol can be adapted to investigate vascular dysfunction and blood-endothelium interactions in other diseases. For complete details on the use and execution of this protocol, please refer to Howard et al.¹.