Tissue engineering and Biomaterials Group, Department of Basic Medical Sciences, Faculty of Medicine and Health Sciences, Ghent University, Ghent, Belgium.
Biofabrication. 2018 Jun 12;10(3):035009. doi: 10.1088/1758-5090/aac7e6.
Overcoming the problem of vascularization remains the main challenge in the field of tissue engineering. As three-dimensional (3D) bioprinting is the rising technique for the fabrication of large tissue constructs, small prevascularized building blocks were generated that can be incorporated throughout a printed construct, answering the need for a microvasculature within the small micron range (<10 μm). Uniform spheroids with an ideal geometry and diameter for bioprinting were formed, using a high-throughput non-adhesive agarose microwell system. Since monoculture spheroids of endothelial cells were unable to remain stable, coculture spheroids combining endothelial cells with fibroblasts and/or adipose tissue derived mesenchymal stem cells (ADSC) as supporting cells, were created. When applying the favorable coculture ratio, viable spheroids were obtained and endothelial cells spontaneously formed a capillary-like network and lumina, as shown by immunohistochemistry and transmission electron microscopy. Especially the presence of ADSC led to a higher vascularization and extracellular matrix production of the microtissue. Moreover, spheroids were able to assemble at random in suspension and in a hydrogel, creating a macrotissue. During at random assembly, cells reorganized, creating a branched capillary-network throughout the entire fused construct by inoculating with capillaries of adjacent spheroids. Combining the advantage of this natural capacity of microtissues to self-assemble and the controlled organization by bioprinting technologies, these prevascularized spheroids can be useful as building blocks for the engineering of large vascularized 3D tissues.
克服血管化问题仍然是组织工程领域的主要挑战。由于三维(3D)生物打印是制造大型组织构建体的新兴技术,因此生成了可整合到打印构建体中的小型预血管化构建体块,满足了在小微米范围内(<10μm)存在微血管的需求。使用高通量非粘附琼脂糖微井系统,形成具有理想几何形状和适合生物打印直径的均匀球体。由于单核细胞培养的内皮细胞球不能保持稳定,因此创建了将内皮细胞与成纤维细胞和/或脂肪组织来源的间充质干细胞(ADSC)作为支持细胞的共培养球体。当应用有利的共培养比例时,获得了有活力的球体,并且内皮细胞自发形成了类似于毛细血管的网络和管腔,如免疫组织化学和透射电子显微镜所示。特别是 ADSC 的存在导致微组织的血管化和细胞外基质产生更高。此外,球体能够在悬浮液和水凝胶中随机组装,形成大组织。在随机组装过程中,细胞重新组织,通过接种相邻球体的毛细血管,在整个融合构建体中创建分支状毛细血管网络。结合微组织自我组装的这种天然能力的优势和生物打印技术的控制组织,这些预血管化球体可用作工程大型血管化 3D 组织的构建体块。
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