Abstract
The nervous system, including the central nervous system and peripheral nervous system, has the most intricate structure and function among all systems in the human body. In studies of physiological and pathological functions, cell culture systems serve as an indispensable tool to simulate the nervous system in vivo. Two-dimensional (2D), three-dimensional (3D), and four-dimensional (4D) neural cell culture systems are used to assess the functional interconnectivity of neuronal tissues and have markedly advanced in recent years. Although 2D culture systems have predominated, they cannot accurately recapitulate the dynamic complexity of the in vivo environment, cell-cell communication, and nervous system structures. Consequently, studies have shifted to using 3D or 4D cell culture systems to achieve more realistic biochemical and biomechanical microenvironments. Nevertheless, many limitations persist in 3D or 4D culture systems, including difficulties in deciphering dynamic and reciprocal remodeling processes, as well as the spatiotemporal distributions of oxygen, nutrients, and metabolic waste. Here, we review 2D, 3D, and 4D culture systems, discuss the advantages and limitations of these techniques in modeling physiologically and pathologically relevant processes, and suggest directions for future research.