Figure 5

Subcutaneous transplantation of ASCVT to nude mice. (a) FbVT and ASCVT were cut out from the Transwell inserts and subcutaneously transplanted to back skin of nude mice. (b–e) Engrafted FbVT (b,c) and ASCVT (d,e) at 2 weeks after the transplantation. (b,d) HE staining; (c,e) immunostaining for human vimentin. The FbVT engrafts as connective tissue containing vascular networks with host blood (b, inset). The thickness of engrafted human-derived tissue is visualized by immunoreaction of human vimentin (c). The ASCVT also subcutaneously engrafted with vascular networks (d), and the thickness was higher than engrafted FbVT (e). (f) Quantitative comparison of the graft thickness between FbVT and ASCVT (N = 7). The ASCVT significantly shows higher thickness. (g–k) High magnified images of engrafted ASCVT. (g) HE staining. The graft contains the capillary- or small venule-like structures with host blood (V) and abundant stroma. (h) MassonGoldner staining for serial section of (g). The stroma stained for collagen fibers, indicating the construction of connective tissue. (i–k) Immunostaining by using antibodies for human CD34 (red) and mouse/human CD31 (green). The engrafted vessels derived from HUVECs are positive for both antibodies (i,j). At the peripheral area of the graft, the vessels with staining by antibodies for mouse/human CD31 and partial staining by antibodies for human CD34 are observed (i,k) suggesting the anastomotic region between host and graft circulation. The nuclei of the cells in the dark field images were visualized by DAPI (blue color). The images are representatives from three independent experiments.