Human tissues and organs have heterogeneous structures with multiple property gradients, which are difficult to restore by single-material bioprinting technology. The advances in multi-material bioprinting technologies have shown great promise in replicating tissue-engineered constructs with one or more functional gradients. In this study, a multi-material 3D printing system based on digital light processing (DLP) was developed, which could efficiently complete multi-material bioprinting tasks. An appropriate concentration of an ultraviolet absorber was selected to improve the printability of channels, and meanwhile, curing parameters were optimized to improve the printing accuracy. The regulation of the mechanical properties of 3D printed constructs was also explored, which offered guidance on the printing of constructs with mechanical anisotropy. In addition, a cell-laden tracheal construct was bioprinted with a biomimetic heterogeneous structure and mechanical gradient, which could support superior cell viability during a 7-day culture. This study showed that the DLP-based process has the capability of building constructs with complex structures and multiple materials, exhibiting the potential to be used in the biofabrication of heterogeneous and functionally-graded tissues and organs for regenerative medicine.