Abstract: The study of central nervous system disease is dependent on in vitro culture of neuronal cells. However, it is hard to determine the interaction between cells in culture of single type of neuronal cells. The co-culture system is able to mimic the cell-cell interaction in the brain and to facilitate investigation into the interaction between different types of cells, as well as cell-environment interaction. The co-culture of neurocytes is more and more popular in the disease study of central nervous system in vitro, such as stroke, Parkinson's disease, Alzheimer's disease, amyotrophic lateral sclerosis, etc. Neurovascular unit (NVU), which is composed of neurons, brain microvascular endothelial cells and astrocytes, can reflect the structure and function of brain in state of the art. Establishment of NVU in vitro is important in the study of the brain diseases. In this paper, several co-culture models of the central nervous system are reviewed in techniques for two-dimensional and three-dimensional culturing. Cell contact and non-contact methods are compared. Moreover, their application in the relevant research and the future direction are explored.