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Fig. 5 | Stem Cell Research & Therapy

Fig. 5

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. 5

Recombinant human uPAR protein promoted engraftment of human myogenic cells in TA muscle of NOD/Scid mice. A Outline of the transplantation experiments. Before transplantation, myogenic cells were cultured in 10% FBS/DMEM supplemented with 20 ng/ml uPAR, BDNF, or IGFBP2 for 3 d. Recombinant uPA was added 24 h before transplantation. The TA muscles of NOD/Scid mice were injured with 1.2% BaCl2 solution 24 h before cell transplantation and injected with 1.5 × 106 cells (Hu5/KD3 cells) or 1.0 × 106 cells (hiPSC-derived muscle progenitors) in 50 μl of PBS containing 5 μg/ml uPAR or BDNF. Two weeks after transplantation, the mice were killed, and the TA muscles were dissected for immunohistochemical analysis using an anti-human lamin A/C antibody and human spectrin (green). B Numbers of human lamin A/C-positive nuclei/section of TA muscle injected with Hu5/KD3 cells treated with the indicated cytokines. n = 4–8 mice/group. Data are shown as the means ± SEMs. Dunnett’s analysis. n.s., not significant. *, p < 0.05. C Hu5/KD3 cells were seeded at a density of 1.0 × 106/well in 6-well collagen plates in the absence or presence of uPAR (10 ng/ml) or uPA (10 ng/ml). The next day, a straight scratch was made with a 200 μl pipette tip (0 h). Migration activity in 5 h was evaluated by measuring the width of the wound. Eight points/condition. Data are shown as the means ± SEMs. Dunnett’s analysis. *, p < 0.05. D Representative immunostaining of cross sections of TA muscles injected with Hu5/KD3 cells with uPAR (5 µg/ml) or uPAR + uPA (5 µg/ml, each). Cells were cultured in 10% FBS/DMEM supplemented with 20 ng/ml uPAR or 20 ng/ml uPAR + 20 ng/ml uPA for 3 d. Myofibers formed by Hu5/KD3 cells were identified with anti-human lamin B1 antibody (red) and human spectrin (green). E Quantitative analysis of (D). n = 4 mice/group. Unpaired two-tailed Student’s t-test

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