Establishment of organ constructing process based on Origami concept

为了验证本研究中基于“折纸”概念的培养组织器官构建过程，构建了具有毛孔结构和血管的完整皮肤。首先，绘制完整皮肤组织发育图。然后，将细胞转移打印技术应用于成纤维细胞、血管细胞、平滑肌细胞的制备，并在具有血管结构的玻璃板上进行血管结构的构建试验。将血管细胞印在成纤维细胞上，毛细血管网状结构轻微形成。此外，将血管细胞印在铺在Vitrigel上的胶原凝胶上，并与胶原凝胶夹在一起，可以有效地形成毛细血管网络结构。另一方面，平滑肌细胞不参与毛细血管网络结构的诱导。此外，构建了包括动脉和静脉在内的结构，以检验培养器官的组装方法，并检查了是否能够产生毛细血管。首先，对含有毛细管网络结构的Vitrigel进行了层合和折叠测试，但无法构建介质的循环系统。此外，由于无法找到有效的黏合剂，在胶原凝胶中构建了模拟血管的微观结构，并与Vtrigel结合，包括毛细血管网络。结果可以进行循环培养，但不能保持毛细管结构。另一方面，尝试制备高功能玻璃体，在玻璃体中加入BMP和肝素。据了解，肝素的加入是无效的，尽管BMP缓释的Vitrigel在小鼠移植实验中促进了骨诱导。

To prove the cultured tissues and organs construction process based on the concept of "Origami" in this research, a complete skin that had the pores structure and the blood vessel was constructed. First of all, the development chart of the complete skin organization was made. Next, the cell transfer printing technology was applied to the fibroblast, the blood vessel cells, and the smooth muscle cell to make it, and the construction of the blood vessel structure was tried on Vitrigel sheet that had the blood vessel structure. When the blood vessel cells were printed on the fibroblast cells, the capillary network structures were slightly formed. In addition, when the blood vessel cells were printed on the collagen gel spread on Vitrigel and sandwiched with collagen gel, the capillary network structures were efficiently formed. On the other hand, the smooth muscle cells did not contribute to the induction of the capillary network structure.Moreover, the structure including the artery and the vein was constructed to examine the method of assembling cultured organs, and whether the capillary was able to be generated was examined. At first, lamination and folding of the Vitrigel including the capillary network structure was tested, but the circulation system of the medium could not be constructed. In addition, because an efficient adhesive agent was not able to be found, the micro structure s mimicking blood vessel were constructed in the collagen gel and were combined with the Vtrigel including the capillary networks. As a result, the circulation culture was able to be done, but, the capillary structure could not be maintained.On the other hand, to attempt making of the high-functional Vitrigel, BMP and Heparin were added in the Vitrigel. It has been understood that the inclusion of Heparin is ineffectual though BMP slow-releasing Vitrigel contributed to the bone inducement as a result of the mouse transplant experiment.