Fig. 3

In vivo Matrigel plug assay of MVFs, SVFs, and MSCs (A) Photographs, H&E staining, and immunostaining of blood vessels of Matrigel plugs. (Top panel) Matrigel solution containing MVFs, SVFs, MSCs, and VEGF was subcutaneously injected, and the representative images of Matrigel plugs collected after 14 days are shown. (Middle panel) Histological analysis of the Matrigel plugs with H&E staining. The newly formed blood vessels containing red blood cells are indicated by arrows. (Bottom panel) Immunostaining of the blood vessels with antibodies against α-SMA and HNA. The nuclei were stained with DAPI, and overlaid images are shown. Scale bar = 100 μm. (B) Quantification of the number of blood vessels in the H&E-stained Matrigel plugs (n = 4). (C) Quantification of hemoglobin levels in the Matrigel plugs using a hemoglobin assay kit (n = 6). (D) Measurement of blood vessel formation by transplanted cells. The numbers of HNA- and α-SMA double positive vessels were counted (n = 4). Data indicate mean ± SD. ^#p < 0.01, *p < 0.05