3D printing technology is widely employed in various scientific disciplines as well as industrial applications such as hearing aid manufacturing. While technological advances and increasing resolution are making 3D printing accessible for microfabrication purposes, one question remains: how can small and delicate components like microgears, lattices, or micromedical devices be released from the build surface of the 3D printer without manual intervention? Herein, a method for 3D printing on top of water-soluble sacrificial substrates made from polyvinyl alcohol (PVA) is presented. Prefabricated sacrificial PVA substrates can be mounted onto a customized holder and serve as a build surface during the 3D printing operation. The substrates do not only facilitate a mild release of 3D printed objects after dissolution of the sacrificial material, they also potentially allow for a convenient manipulation and further array-based processing of predetermined patterns of printed structures subsequent to the 3D printing procedure. This, in turn, may enable a full integration into automated production lines. The fabrication of PVA substrates is thoroughly characterized and the 3D printing of various exemplary structures on sacrificial substrates is demonstrated. Finally, the release of 3D printed objects from PVA substrates is shown.