Fig. 5

The hESC-MSCs could significantly improve the function of ischemic hind limbs in vivo. a Percentage distributions of limb loss, foot necrosis, and limb salvage were analyzed by Fisher’s exact test. b Semi-quantitative clinical assessments of ambulatory impairment (left) and ischemic damage (right) of ischemia hind limbs among the Ctr, +PBS, +hESC-MSC, and hBM-MSC groups. Data are shown as mean ± SEM (n = 6). *P < 0.05; **P < 0.01. c In vivo dynamically monitoring the status of angiogenesis after transplantation of hESC-MSCs or hBM-MSCs through tracking Vegfr2-luc expression by BMI in hind limb ischemic models at day 21. d Dynamic tendencies of angiogenesis in hind limbs based on Fluc signals among the indicated groups. e In vivo monitoring the retention of hESC-MSCs or hBM-MSCs by BLI analysis. f Representative images of hind limb muscle sections stained for CD31 (red) at day 14. DAPI was used for nucleus staining. g Quantification of capillary density of ischemic hind limbs in the indicated groups. Data are shown as mean ± SEM (N = 3). *P < 0.05; ^#P < 0.05. h Histopathologic analysis of muscle tissues of mice (Ctr, +PBS, +hESC-MSCs) at day 28 by hematoxylin and eosin (H&E) staining (scale bar = 100 μm). i Histopathologic analysis of muscle tissues of mice (Ctr, +PBS, +hESC-MSCs) at day 28 by Masson staining (scale bar = 100 μm). j Quantitative analysis of collagen content in fibrotic area with Masson staining (Ctr, +PBS, +hESC-MSCs). Data are shown as mean ± SEM (n = 6). **P < 0.01. k Typical morphology of hESC-MSCs or hBM-MSCs secreted exosome under transmission electron microscopy. Scale bar, 200 nm. l qRT-PCR analysis of the cytokines associated with angiogenesis (VEGFA, VEGFB, ANG1, ANG2) in HUVECs after culturing with hESC-MSCs and hBM-MSCs for 2 days. Data are shown as mean ± SEM (n = 3). *P < 0.05; **P < 0.01. All values are normalized to the NC group (= 1)