Fig. 2.

The SHH pathway enhances angiogenic potential of WJ-MSC in vitro. Serum-starved HUVEC, seeded in Matrigel, were treated with conditioned medium (CM) from nonstimulated WJ-MSC or with CM from purmorphamine (Pur)-stimulated WJ-MSC (CM + Pur) at the indicated time points. EGM-2 was used as positive control (Ctl +) and DMEM as negative control (Ctl -). The angiogenic response was assessed by counting the number of branches and tubules formed by endothelial cells after 3–4 h. a–f Representative images of different treatments. g, h Histograms comparing the number of branching points and tubules between control and experimental conditions as indicated. While there is no difference in these parameters between CM obtained after 24 and 48 h under standard conditions, a strong response was observed after 48 h CM + Pur treatment (CM obtained from 48-h Pur-treated WJ-MSC) which indicates that SHH pathway stimulation enhances the angiogenic capacity of WJ-MSC. CM and CM + Pur were obtained from three independent WJ-MSC cultures. ***P < 0.0001, one-way ANOVA. Scale bar = 100 μm. ns not significant