There is an increasing prevalence of neuropathologies resulting from traumatic brain injury, as well as neurodegenerative diseases (ND) such as Alzheimer’s, Parkinson’s, Huntington’s, particularly among elderly patients. The nervous system regeneration necessitates the repair or replacement of damaged nerve cells which, however, exhibit limited renewal capacity, and the complex nature of NDs hinders the development of effective treatments. Limitations of standard 2D cell cultures, as well as in vivo animal models, can be overcome using 3D cultures and co-cultures such as cell spheroids, organoids and organs-on-chip, by enabling the study of cell-cell and cell–extracellular matrix (ECM) interaction and replication of brain functionality.