Cryobioprinting for Shelf-Ready Tissue Fabrication and Storage

摘要 三维生物打印为制造复杂的细胞组织铺平了一条新的途径 构造。到目前为止，生物打印已被广泛应用于组织工程、组织工程、组织工程等领域 模型工程、有机化合物生产等。然而，存在着关于 可供货架使用的3D生物打印组织构造物的制造和储存。由于涉及的内在复杂性 在大多数生物印刷过程中，包括广泛采用的挤压生物印刷，使用这种方法作为一种 在许多情况下，现场制作技术可能很繁琐，有时甚至不切实际。 此外，缺乏一种长期储存细胞组织结构的功能性方法排除了 预制生物印刷产品的货架供应情况。在这里，我们建议开发一种独特的低温生物打印 通过无缝方式同时制造和存储细胞负载的体积组织结构的策略 结合细胞挤压法、生物印迹法和超低温保存法。低温生物打印的性能将是 通过设计、制造和存储由低温保护性生物墨水制成的细胞加载结构来进行研究 冷冻板，温度精确可控。预计原位冻结过程将进一步 促进细胞水凝胶生物墨水的印刷适宜性，以实现自由结构，否则通常 直接挤压生物印花不方便。我们相信，我们的低温生物打印将一步到位 一种并行组织生物制造和存储的方法。

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.