Studying neurological disorders in vitro is still challenging due to the human brain’s complexity and the difficulty of obtaining primary neural cells. However, tissue engineering and tridimensional (3D) cell culture have become increasingly important tools for disease modeling. By providing an extracellular matrix (ECM) substrate that closely resembles physiological conditions, 3D cell culture offers several advantages over standard monolayer cell culture, including enhanced cell-cell and cell-matrix interactions. This results in a microenvironment that more accurately reflects in vivo biology. In this study, we performed an in-depth analysis of the proteome and matrisome of 3D tissue-engineered dermis, made from human primary dermal fibroblasts cultivated in 3D and embedded in a self-produced ECM. Interestingly, in silico analysis revealed that neurogenesis and associated functions were predicted to be strongly activated in this tissue-engineered 3D model. Indeed, we showed that ECM proteins involved in neuronal development and maintenance, typically produced by cerebral cells, were also expressed by dermal fibroblasts. Of particular interest, the 3D co-cultivation of dermal fibroblasts with iPSC-derived motor neurons readily enabled long-lasting culture periods without costly media supplementation with exogenous additives. Patient-derived dermal fibroblasts, cultivated in 3D, could therefore become valuable models for the study of neurological diseases. This approach offers a cost-effective and a less invasive alternative to brain biopsies for modeling complex neurological disorders in vitro.