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Fig. 4 | Stem Cell Research & Therapy

Fig. 4

From: Mesenchymal stem cells derived from human induced pluripotent stem cells improve the engraftment of myogenic cells by secreting urokinase-type plasminogen activator receptor (uPAR)

Fig. 4

uPAR and BDNF stimulated migration of Hu5/KD3 cells in vitro. A Hu5/KD3 cells were seeded at a density of 1.0 × 106/well in 6-well collagen plates in the presence of uPAR (left), BDNF (middle), or IGFBP2 (light) (0–50 ng/ml). The next day, a straight scratch was made with a 200 μl pipette tip (0 h) and photographed under an inverted microscope. Six hours later, the cells were again photographed (Additional file 1: Fig. 6). Six views/condition were recorded. Migration activity was evaluated by measuring the width of the wound. B Hu5/KD3 cells were plated onto 24-well collagen plates at 2 × 105 cells/well and induced to differentiate in 10% FBS/DMEM medium supplemented with recombinant human uPAR (left), BDNF (middle) or human IGFBP2 protein (right) (0–50 ng/ml). Four (uPAR) or five days (BDNF, IGFBP2) later, the cells were fixed and stained with an anti-muscle myosin heavy chain antibody (MF20) and DAPI (nuclei). Shown are averages of 4 wells/condition. The percentage of MF20-positive area was quantified using ImageJ software. C Hu5/KD3 cells were plated onto 24-well collagen plates at 1 × 104 cells/well in 10% FBS/DMEM supplemented with recombinant human uPAR (left), BDNF (middle), or IGFBP2 (right) protein (0–50 ng/ml). Three days later, an MTT assay was performed (4 wells/condition). In (A–C), data are shown as the mean ± SEM. Dunnett’s analysis. n.s., not significant. **, p < 0.01. ***, p < 0.001. ****, p < 0.0001

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