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A 3D cell culture has developed rapidly in recent years, as cells growing on a flat substrate in a static environment are far from achieving an in vivo status. Currently, researchers have also gradually realized that to achieve cell morphology, structure, and physiological functions in vitro, 3D cell culture should be capable of simulating key features of an in vivo environment, including the interaction of cell–cell, cell–extracellular matrix (ECM), and cell–organ interactions. Herein, the development of the 3D cell culture system related to the following three perspectives is outlined: 1) biomaterial systems with a hydrogel system as the core; 2) biomanufacturing technology with bioprinting as the main means; and 3) culture device systems supported by microfluidic chips and bioreactors. The question is whether 3D cell culture will be as popular as 2D culture in the future. The key may lie in the development of simple and standard protocols for 3D culture.