Figure 3

Perivascular differentiation of hASCs in ASCVT. (a) Highly magnified image of epon-embedded and sectioned ASCVT with toluidine blue staining. V vascular structures. Perivascularly localized hASCs are observed (red arrowheads). (b–g) Immunostaining for human CD34 (hCD34)/human CD90 (hCD90) (b,c,f) and human CD31 (hCD31)/αSMA (d–g). (b–e) ASCVT. Vascular structures (V) are immunostained for hCD34 and hCD31. Perivascularly localized hASCs show intense immunostaining for hCD90 and αSMA (c,e). (f,g) Artificial tissue constructed by hASCs without HUVECs. No vascular structures are found. A weak staining of hCD90 and αSMA is observed. (h,i) Perivascularly localized hASCs showing immunostaining for NG2 (yellow arrowheads). (j) Perivascularly localized hASCs showing immunostaining for desmin (yellow arrowheads). (k–m) Addition of EGFP-labeled hASCs to fibroblast-based vascular tissue (NHDFs:hASCs:HUVECs = 7:1:1). (k) Immunostaining of the paraffin section. hASCs with EGFP localized to perivascular area (yellow arrowheads). (l) Highly magnified image of hatched box in (k). EGFP-positive hASC connected to the vascular structure (V). (m) 3D-constructed confocal image. EGFP-positive hASC surrounded the vascular structure like a pericyte. The nuclei of the cells were visualized by DAPI (blue color). The images are representatives from two independent experiments.