3D bioprinting is an emerging manufacturing approach to fabricate (cell-laden) hydrogel constructs with embedded microchannels, which are potentially useful for fundamental studies to understand vascularization and angiogenesis, and for developing organ-on-a-chip devices for disease modeling. Although numerous printing approaches have been developed, novel approaches are still needed that enable printing of channels with user-defined and tunable size, morphology, and complexity. Here, we report a novel bioprinting approach enabling printing of a sacrificial ink within commonly used photocurable hydrogels using a sequential printing approach. To achieve this, photocurable hydrogel is printed layer-by-layer as usual, but each layer is exposed to light briefly (seconds) to create partially crosslinked, self-supporting layers. At a desired thickness, immediately after the layer is printed (prior to partial crosslinking step), sacrificial hydrogel is directly printed within this viscous uncrosslinked layer. The layer was then exposed to light to confine and support the sacrificial hydrogel. After fully crosslinking the system, the sacrificial hydrogel is washed away, forming a channel. This approach allows bioprinting of cells with the matrix material and seeding of cells into channels after the sacrificial ink is removed. This approach can potentially provide a robust platform for fabricating vascularized tissues and studying cell behaviors on diverse channel surfaces. STATEMENT OF SIGNIFICANCE: 3D bioprinting is an emerging platform for the fabrication of hydrogel-based constructs for in vitro tissue/disease modelling or tissue and organ printing. Although several approaches have been developed to print channels within these constructs, it is still challenging to incorporate microchannels (for vascularization) within 3D bioprinted constructs. This study presents a novel bioprinting approach to create user-defined and tunable channels embedded within cell-laden hydrogel constructs. We report an important advance as our approach does not require complex device modifications for bioprinters or complex synthesis and processing hurdles for the inks. Since our approach does not require special chemistries, there are potentially a greater number of commercially available options for ink materials.,Copyright © 2019. Published by Elsevier Ltd.