Dow Silicones Belgium SRL 1 article published in JoVE Bioengineering Cultivating a Three-dimensional Reconstructed Human Epidermis at a Large Scale Irini M. Dijkhoff1, Benedetta Petracca1,2, Roxane Prieux3, Giuseppe Valacchi3,4, Barbara Rothen-Rutishauser1, Marc Eeman2 1Adolphe Merkle Institute, University of Fribourg, 2Dow Silicones Belgium SRL, 3Department of Biomedical and Specialist Surgical Sciences, University of Ferrara, 4Department of Animal Sciences, Plants for Human Health Institute, North Carolina State University This protocol describes a straightforward method to cultivate three-dimensional reconstituted human epidermis in a reproducible and robust manner. Additionally, it characterizes the structure-function relationship of the epidermal barrier model. The biological responses of the reconstituted human epidermis upon proinflammatory stimuli are also presented.