Establishment of organ constructing process based on Origami concept

为了验证本研究中基于“折纸”概念的培养组织和器官构建过程，构建了具有孔隙结构和血管的完整皮肤。首先绘制了完整皮肤组织的发育图。接着，将细胞转印技术应用于成纤维细胞、血管细胞和平滑肌细胞来制造，并且在具有血管结构的Vitrigel片材上尝试血管结构的构建。当血管细胞被打印在成纤维细胞上时，毛细血管网络结构轻微形成。此外，当将血管细胞打印在涂在Vitrigel上的胶原凝胶上并夹有胶原凝胶时，有效地形成了毛细血管网络结构。另一方面，平滑肌细胞对毛细血管网结构的诱导没有贡献。此外，构建了包括动脉和静脉的结构，以检查组装培养器官的方法，并检查是否能够产生毛细血管。首先，测试了包括毛细网络结构的Vitrigel的层压和折叠，但不能构建培养基的循环系统。此外，由于无法找到有效的粘合剂，因此在胶原凝胶中构建了模拟血管的微结构，并与包括毛细血管网络的Vtrigel结合。另一方面，为了尝试制作高功能的Vitrigel，在Vitrigel中加入BMP和肝素。据了解，肝素的加入是无效的，尽管BMP缓释Vitrigel作为小鼠移植实验的结果有助于骨诱导。

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.