Figure 5.
MSX1+PDGFRAlow LM-like cells exhibited strong osteochondral competence
(A) Schematic illustration for the strategy to characterize the osteochondral competence of hPSC-derived LM-like cells.
(B) Confirmation of osteogenic and chondrogenic capability of LPM- and LM-like cells in vitro. Stronger staining of alizarin red and Alcian blue was seen in LM-like cells (MSX1+PDGFRAlow) in comparison with LPM-like ones (MSX1+PDGFRA+), suggesting the enhanced osteochondral potential of the LM-like cells. Scale bars: 100 μm.
(C) Quantitative analysis of the alizarin red (top) and Alcian blue (bottom) staining. Msx1+PDGFRAlow cells had better potential to form more mineralized matrix and larger chondrospheres. Top: error bars represent data from six samples in three independent experiments (mean ± SD). Bottom: error bars represent data from twelve chondrospheres from three independent experiments (mean ± SD). Statistics: independent-sample t test using SPSS version 22.0. ∗∗∗p < 0.001.
(D) Efficient regeneration of articular cartilage by LM-like cells in vivo. Representative images are shown. Alcian blue and COL II staining (black dashed line) were used to detect cartilage formation. Scale bars: 250 μm.
(E) Quantitative analysis of the regenerated cartilage. Alcian blue staining was performed in 3 week samples. Error bars represent data from twelve samples of three independent experiments (mean ± SD). Statistics: independent-sample t test using SPSS version 22.0. ∗∗∗p < 0.001.
(F) Confirmation of human origin in the transplanted cells. Representative images of KU80 immunostaining (white dashed line) in the articular cartilage defect sites are shown. Scale bars: 250 μm.
(G) Repaired articular cartilages were maintained after 8 weeks. Alcian blue and COL II staining (black dashed line) were used to detect cartilage formation. Scale bars: 250 μm.
(H) Quantitative analysis of the regenerated cartilage. Error bars represent data from twelve samples of three independent experiments (mean ± SD). Statistics: independent-sample t test using SPSS version 22.0. ∗∗∗p < 0.001.