Microphysiological systems (MPS) are advanced platforms that mimic the functions of human tissues and organs, aiding in drug development and disease modeling. Traditional MPS fabrication mainly depends on silicon-based microfabrication techniques, which are complex, time-consuming, and costly. In contrast, 3D printing technologies have emerged as a promising alternative, allowing for the rapid and precise creation of intricate three-dimensional structures, thereby opening new avenues for MPS research. This review examines the principles, characteristics, advantages, and limitations of key 3D printing techniques, including fused deposition modeling (FDM), stereolithography (SLA)/digital light processing (DLP), inkjet 3D printing, extrusion-based bioprinting, and laser-assisted bioprinting. Additionally, we discuss how these technologies are applied in MPS fabrication and their impact on MPS research, along with future prospects for advancements in the field.