Fig. 8

In vivo characterization of pre-vascularized DESS containing CD146⁺ pericytes. A The hematoxylin–eosin (H/E) staining of pre-vascularized DESS after 1 week shows a stratified epidermis and dermis containing fibroblasts. Scale bar: 100 μm. B The immunofluorescence staining for CK10 displays a positive signal of this keratinocyte differentiation marker from the suprabasal layer. Laminin332 and CD90 staining indicates the presence of basement membrane and human dermal fibroblasts, respectively. Scale bar: 100 μm. C The immunofluorescence staining for CK19 shows the presence of CK19-positive keratinocytes in both the basal and suprabasal layers. The staining for Laminin332 indicate the presence of the basement membrane. Scale bar: 100 μm. D, E Human capillaries positive for the specific human CD31 antibodies are distributed throughout the human dermis, which is marked with human CD90. Inosculation between human and rat capillaries is visualized by co-localization of humanCD31 and ratCD31 (D, E, insets). The ingrowth of rat capillaries is also observed in the human neo-dermis (arrows). White dashed lines indicate the dermo-epidermal junction. Scale bar: 100 μm, inset 50 μm. F The green autofluorescence of rat erythrocytes (ratBCs) inside the lumen of human capillaries indicates the presence of perfused capillaries. Scale bar: 100 μm, inset 50 μm. G Immunofluorescence staining for CD146 and humanCD31 shows the co-localization between these two markers and the presence of human CD146⁺ pericytes around capillaries. Scale bar: 100 μm, inset 50 μm. H Lymphatic capillaries stained for Lyve1 are CD146 negative. I Human pericytes double-positive for CD146 and NG2 are observed around capillaries. CD146 negative capillaries, attributable to lymphatic capillaries, show no presence of pericytes (asterisk). G, H, I (n = 3 independent donors). White dashed lines indicate the epidermal-dermal junction. Scale bar: 100 μm, inset 50 μm