Please cite this article in press as: Teixeira, D.T.D. et al. Bioengineered cell culture systems of central nervous system injury and disease, Drug Discov Today (2016), http://dx.doi.org/ 10.1016/j.drudis.2016.04.020 Drug Discovery Today  Volume 00, Number 00  May 2016 REVIEWS Bioengineered cell culture systems of central nervous system injury and disease Fa ´ bio G. Teixeira 1,2 , Nata ´ lia L. Vasconcelos 1,2 , Eduardo D. Gomes 1,2 , Fernanda Marques 1,2 , Joa ˜o C. Sousa 1,2 , Nuno Sousa 1,2 , Nuno A. Silva 1,2 , Rita Assunc ¸a ˜ o-Silva 1,2 , Rui Lima 1,2 and Anto ´ nio J. Salgado 1,2 1 Life and Health Sciences Research Institute (ICVS), School of Health Sciences, University of Minho, Braga, Portugal 2 ICVS/3B’s, PT Government Associate Laboratory, Braga/Guimara ˜es, Portugal Cell culture systems, either 2D or explant based, have been pivotal to better understand the pathophysiology of several central nervous system (CNS) disorders. Recently, bioengineered cell culture systems have been proposed as an alternative to the traditional setups. These innovative systems often combine different cell populations in 3D environments that more closely recapitulate the different niches that exist within the developing or adult CNS. Given the importance of such systems for the future of CNS-related research, we discuss here the most recent advances in the field, particularly those dealing with neurodegeneration, neurodevelopmental disorders, and trauma. Introduction Injury and disease within the CNS frequently induce chronic and acute insults leading to irreversible processes resulting in neuronal cell death. This fact is intimately related to the low regenerative potential of the CNS and the complexity of its several niches. Additionally, the causes that induce such phenomena are multivar- iate in nature. Indeed, cell death within the CNS can be triggered by injury, as in traumatic brain injury (TBI) or spinal cord injury (SCI); protein aggregation, such as in the case of Alzheimer’s (AD) or Parkinson’s disease (PD); neurodevelopmental-related problems, such as Rett syndrome; or induced neurodevelopmental neurotox- icity (DNT) phenomena. Thus, understanding the mechanisms behind such pathologies, as well as the possible therapeutic strate- gies that could be used to counteract them, is essential. To do so, it is essential to understand how CNS cells operate under these condi- tions, as well as how they interact with each other. Animal models of injury and disease, as well as CNS cell culture systems, are often use to assess CNS cell interactions and how they operate. Models of injury are often seen as the last phase of preclinical research and try to replicate, as far as possible, the molecular, biochemical, and phenotypical characteristics of the CNS condition under study. However, for the initial screening of therapies, such as a library of small molecules with potential therapeutic use, such models might not be the best option. For instance, the differences between animal and human cell biology can lead to misleading results in important areas, such as toxicol- ogy [1,2]. Models of disease have been also used for years to understand the molecular mechanisms of injury and disease of the CNS, as well as for the early development of therapeutic strategies. Within these models, cell lines such as SH-SY5Y [3], N2 [4] or PC-12 [5], or MO-4 [6] are commonly the first system to be used. However the gold standard within the field are primary cultures of CNS cells [neural stem cells (NSC), neurons, astrocytes, oligodendrocytes, and microglial cells], which are often isolated from embryonic or early postnatal [up to postnatal day (P)5] tissue from rodents, mainly mice and rats [7]. Other popular systems within the field are explant cultures [8] and organotypic-like cultures [9–11]. Explant cultures are often used to study, for example, the development of axons and processes associated with it, whereas organotypic-like cultures are popular in the study of how CNS cells work in a 3D environment, such as the spinal cord [9], hippocampus [10] or substantia nigra/striatum [11]. Reviews  POST SCREEN Corresponding author: Salgado, A.J. (asalgado@ecsaude.uminho.pt) 1359-6446/ß 2016 Published by Elsevier Ltd. http://dx.doi.org/10.1016/j.drudis.2016.04.020 www.drugdiscoverytoday.com 1