Study of neurological pathology mechanisms is essential to understand and create therapies of neurological disorders. However, due to the human brain’s complexity and inaccessibility, research is limited. Additionally, there isn’t a well established neuronal cell line available and animal studies fail to reflect physiology of human brain disorders. To address these limitations, stem cell technologies are being developed to create more accurate disease models. Mesenchymal stem cells have emerged as a preferred option for neuronal differentiation due to their multipotency, accessibility, and low immune response properties. Various methods have been employed to differentiate mesenchymal stem cells into neurons, but it is crucial to ensure that the resulting neurons possess functional characteristics resembling a complete neuronal unit within the model. In this study, we describe the process of differentiating human bone marrow-derived mesenchymal stem cells into neurons using a specific differentiation medium supported with extracellular matrix components. We evaluate the functionality of these differentiated neurons using multielectrode arrays, providing possible valuable insights for future research in this field.