Although various types of organs-on-chips have been introduced recently as tools for drug discovery, the current studies are limited in terms of fabrication methods. The fabrication methods currently available not only need a secondary cell-seeding process and result in severe protein absorption due to the material used, but also have difficulties in providing various cell types and extracellular matrix (ECM) environments for spatial heterogeneity in the organs-on-chips. Therefore, in this research, we introduce a novel 3D bioprinting method for organ-on-a-chip applications. With our novel 3D bioprinting method, it was possible to prepare an organ-on-a-chip in a simple one-step fabrication process. Furthermore, protein absorption on the printed platform was very low, which will lead to accurate measurement of metabolism and drug sensitivity. Moreover, heterotypic cell types and biomaterials were successfully used and positioned at the desired position for various organ-on-a-chip applications, which will promote full mimicry of the natural conditions of the organs. The liver organ was selected for the evaluation of the developed method, and liver function was shown to be significantly enhanced on the liver-on-a-chip, which was prepared by 3D bioprinting. Consequently, the results demonstrate that the suggested 3D bioprinting method is easier and more versatile for production of organs-on-chips.