Colony Formation, Migratory, and Differentiation Characteristics of Multipotential Stromal Cells (MSCs) from “Clinically Accessible” Human Periosteum Compared to Donor-Matched Bone Marrow MSCs

Figure 5

Trilineage differentiation assays, comparing donor-matched periosteum and bone marrow MSCs. Each assay was assessed using (a) histology staining, (b) quantitative assays, and (c) qPCR for specific markers of differentiation. Osteogenic assays stained for alkaline phosphatase (fast blue) and calcium deposition (alizarin red), with quantified Ca²⁺ content and assessment of runt-related transcription factor 2 (RUNX2) and bone gamma-carboxyglutamate (gla) protein (BGLAP) expression levels. Chondrogenic assay macro images (insert) stained for glycosaminoglycans (GAG) (toluidine blue), quantified GAG content and collagen, type 2, alpha 1 (COL2A1), and SRY- (sex-determining region Y-) box 9 (Sox9) expression levels. Adipogenic assays stained for fat deposition (oil red), quantified fat deposition (Nile red) and nuclei content (DAPI), and fatty acid-binding protein 4 (FABP4) and peroxisome proliferator-activated receptor-gamma (PPAR-γ) expression levels. Wilcoxon signed-rank test was carried out, . Scale bars represent 500 μm (fast blue, alizarin red, and oil red), 200 μm (macro insert), and 200 μm (toluidine blue). BM: bone marrow, P: periosteum, bd: below detection.

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