Cryobioprinting for Shelf-Ready Tissue Fabrication and Storage

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Abstract Three-dimensional (3D) bioprinting has paved a new avenue for fabricating intricate cell-laden tissue constructs. So far, bioprinting has been adopted in myriad applications such as tissue engineering, tissue model engineering, and organoid production, among others. There are, however, challenges regarding the fabrication and storage of shelf-ready 3D-bioprinted tissue constructs. Due to the intrinsic complexities involved in most bioprinting processes, including the broadly adopted extrusion bioprinting, using this method as an on- site fabrication technique can be cumbersome, or sometimes impractical under a number of scenarios. Furthermore, the lack of a functional approach for long-term storage of cell-laden tissue constructs precludes the shelf-availability of pre-made bioprinted products. Here, we propose to develop a unique cryobioprinting strategy for simultaneously fabricating and storing cell-laden volumetric tissue constructs through seamlessly combining cell-laden extrusion bioprinting and cryopreservation. The cryobioprinting performances will be investigated by designing, fabricating, and storing cell-laden constructs made of cryoprotective bioinks using a freezing plate with precisely controllable temperature. The in situ freezing process is further anticipated to promote the printability of cell-laden hydrogel bioinks to achieve freeform structures otherwise oftentimes inconvenient with direct extrusion bioprinting. We believe that our cryobioprinting will emerge as a single-step method for concurrent tissue biofabrication and storage.