Fig. 5: Neural crest-derived multipotent cells contribute to regenerated joint tissues.
a, Lineage trace of DsRed-labeled neural crest lineage-derived cells using Sox10:Cre;actb2:loxP-BFP-STOP-loxP-DsRed (n = 3-5 per timepoint); bone visualized with calcein green staining. White arrow denotes regenerated joint surface and white arrowhead indicates regenerating chondrocytes. White dashed lines indicate cut sites. b, Simplex plots of archetype analysis colored by cluster and split by time point. c, UMAP of skeletal cells colored by most similar archetypes. d, Distribution of pseudotime of skeletal cell clusters across timepoints. The center of each box plot is marked by a horizontal line and represents the median. The bounds of the box are specified by the lower quartile (25th percentile) and the upper quartile (75th percentile). The whiskers extend from the box and denote the data points that fall within 1.5 times the interquartile range. e, Cell type composition of cells at 1 dpjr that have pseudotime between 0 and 2. f, Simplex plots of archetype analysis with osteoblasts, chondrocytes, chondrogenic fibroblasts, and regenerative mesenchyme colored by pseudotime. g, Single-clone lineage trace of regenerated Zebrabow ligament from Sample 1 (n = 5). Orange arrow denotes green ligamentocyte circled in the corresponding polar graph. h, Single-clone lineage trace of regenerated Zebrabow joint from Sample 1 (n = 5). Orange arrowhead denotes green chondrocyte circled in the corresponding polar graph. i, Polar graph of HS color distribution of regenerated cells from Sample 1. Orange circle indicates a green ligamentocyte and chondrocyte clone pair fewer than 10 units apart in color space. j, Histogram displaying the frequency distribution of distances between regenerated cells from Sample 1 in HS color space. Approximately 10% of cell pairs in Sample 1 are predicted clones. k, Single-clone lineage trace of regenerated joint tissues from Sample 2 (n = 5). Orange arrowhead denotes cyan chondrocyte circled in the corresponding polar graph. Orange arrow denotes cyan adipocyte circled in the corresponding polar graph. l, Polar graph of HS color distribution of regenerated cells from Sample 2. Orange circle indicates a cyan adipocyte and chondrocyte clone pair fewer than 10 units apart in color space. m, Histogram displaying the frequency distribution of distances between regenerated cells from Sample 2 in HS color space. Approximately 17.5% of cell pairs in Sample 2 are predicted clones. Scale bars, 250 µm (a), 125 µm (g,h,k). AA, anguloarticular bone; IOP, interopercular bone; Q, quadrate bone; RA, retroarticular bone.
