Infection with herpes simplex virus type 1 (HSV-1) causes the most common skin disease. Various test systems have been established to recapitulate this cyclical pathway of productive infection, latency, and reactivation. Most studies of latency and reactivation are conducted in animal models. However, the small number of neurons which harbor the viral genome, the complexity of the in vivo setting, and ethical constraints place limits on animal studies. So far, no in vitro model which resembles natural latency exists. Here, we describe the first in vitro HSV-1 infection model based on a human skin equivalent. The 3D infection model is generated using the human keratinocyte cell line HaCaT grown on a collagen substrate containing human primary fibroblasts and in addition a quiescently HSV-1 infected neuronal component.