Each year, snakebite envenoming claims 100,000 lives and leaves 400,000 disabled victims behind. Many snakebite victims survive the lethality of snake venom, yet suffer from local damages it may cause, such as chronic ulcers and malignant transformations. These complications are usually caused by locally acting toxins, such as cytotoxins, which bind to different cells, particularly in skin and muscles, and disrupt the cellular membranes. The wound that results from the action of these toxins may, in turn, evolve into extensive necrosis, which may necessitate debridement, grafting, or, in more severe cases, amputation. In this relation, it is estimated that at least 6,000 snakebite-related amputations occur on a yearly basis in Africa alone1. Currently, murine models of envenoming are the golden in vivo standard when studying venom-induced dermonecrosis2. However, even if ethical considerations are set aside, data obtained from animal studies might not faithfully mimic the pathophysiological effects seen in humans. In order to assess the mechanisms by which snake toxins affect humans, a physiologically more relevant, and technologically more advanced alternative to animal models is the use of 3D organotypic models of human cells which, in comparison to monolayer cell cultures, replicate cell composition complexity of tissues and spatial cell-cell or cell-extracellular matrix interactions in a more effective manner3. Here, we report the establishment of the first 3D organotypic model of human skin for investigating the dermonecrotic effects of Naja nigricollis envenoming. It is anticipated that this study can help deepen our understanding of the mechanisms underlying local necrosis induced by snakebite envenoming in humans and thereby aid the development of better envenoming therapies, including next-generation recombinant antivenoms, to reduce the burden of local tissue damage and amputations amongst snakebite victims.