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. 2022 Sep 5;13:5211. doi: 10.1038/s41467-022-32868-y

Fig. 6. In situ expansion of Msx1+ SSCs subset promoted efficient bone regeneration partially through endochondral ossification.

Fig. 6

a Trajectory of differentiation from cycling MSCs to both SSC1 and SSC2 lineages predicted by Monocle 2. b Heatmap of gene expressions in subsets ordered by pseudotime of two differentiation trajectories in a. c Distribution of cells on both the differentiation trajectories from Defect and NSs groups showing a featured change dominated at 1 week and 2 weeks after surgery, respectively. d Relative expression level of anabolic and metabolic genes (Acan, Col6a5, Mmp13) of cartilage and Msx1 gene along the whole pseudotime. e Expression of the above chondrocyte-specific genes (Acan, Col6a5, Mmp13) and SSC2 marker gene (Msx1) visualized on differentiation trajectory. f Co-immunostaining of Msx1 and Acan expression of paraffin sections in Scaffold + NSs group at 1 week and 2 weeks after defect surgery, respectively (bar = 200 μm at low magnification and bar=30μm at high magnification). g Co-immunostaining of Msx1 and Mmp13 expression of paraffin sections in Scaffold + NSs group at 1 week and 2 weeks after defect surgery, respectively (bar = 200 μm at low magnification and bar = 30 μm at high magnification). h, i Safranin-O staining of paraffin sections in Untreated Defect group and Scaffold + NSs group at 1 week and 2 weeks after defect surgery, respectively (bar = 500 μm at low magnification and bar=50μm at high magnification). At least three times of experiments were repeated independently.