Protocol for designing and bioprinting multi-layered constructs to reconstruct an endothelial-epithelial 3D model

3D bioprinting has opened new possibilities and elevated tissue engineering complexity. Here, we present a protocol to design a 3D model with two cell lineage layers (A549 and HUVEC) to recreate multi-cell constructs. We describe the steps for slicing the constructs, handling hydrogels, and detailing the bioprinting setup. These 3D-bioprinted constructs can be adapted to various cell models—from primary cell cultures to commercial cell lines and induced pluripotent stem cells (IPCs)—and applications, including drug screening and disease modeling. For complete details on the use and execution of this protocol, please refer to Cruz et al.(1)