Table 1.
Cell compositions of spheroids.
Fig 1.
Cells: fibroblasts, mesenchymal stem cells, smooth muscle cells, or endothelial cells were cultured respectively. Multicellular spheroids were created using mixed cell suspensions, and the artificial esophagus was then constructed with bio-3D printing using those spheroids. The artificial esophagus was matured in a bioreactor for a total of 4 weeks. Finally, esophageal transplantation of the artificial esophagus was performed.
Table 2.
Results of the tensile test in the bio-3D printed structures and the native esophagus of rats.
Fig 2.
Mechanical characteristics of structures made by the bio-3D printer.
A. Maximum load tensile elongation B. Tensile strength The mechanical data of the structures measured by a Tissue Puller.
Fig 3.
Macroscopic and immunohistochemical findings of pre-transplanted structures.
The bio-3D printed structures are shown before transplantation. A-D: HE staining. E-H: Immunohistochemical staining with anti-αSMA antibodies. I-L: Immunohistochemical staining with anti-VEGF antibodies. M-P: immuno-histochemical staining with anti-CD31 antibodies. Q-T: Immunohistochemical staining with anti-elastin antibodies. Scale bar = 100 μm. HE: hematoxylin-eosin, SMA: smooth muscle actin, VEGF: vascular endothelial growth factor.
Fig 4.
Transplantation of the structures.
A. The surgical site of transplantation. B. The transplanted site at 30 days after transplantation. Arrows: transplanted site, Du: duodenum, E: esophagus, EG: esophagogastric junction, L: liver, St: stomach.
Fig 5.
Assessment of the structures at 30 days after transplantation.
A: The transplanted site. B: Hematoxylin and eosin (HE) staining at the site shown in A (yellow box). Scale bar = 500 μm. C-E: HE staining of the transplanted structure. Scale bar = 500 μm (C), 100 μm (D), and 50 μm (E). F-H: Immunohistochemical staining of the transplanted structure with anti-pan-cytokeratin. Scale bar = 500 μm (F), 100 μm (G), and 50 μm (H). I-K: Immunohistochemical staining of transplanted structure with anti-αSMA. Scale bar = 500 μm (I), 100 μm(J), and 50 μm (K). Arrows: transplanted structure, E: esophagus, St: stomach. Immunohistochemical staining of the transplanted structure with anti-HLA class 1 ABC. Scale bar = 200 μm (L), 100 μm (M).