Characterization of Mesenchymal Stem Cell-Like Cells Derived From Human iPSCs via Neural Crest Development and Their Application for Osteochondral Repair

Figure 4

Chondrogenesis of iNCMSCs. The representative data from three lines is shown. (a) Safranin-O staining of chondrogenic pellet stimulated with TGFβ3 and BMP2 at day 28. High magnification image is shown in the right panel with the same frame. (b, c) Immunostaining for COL2 and SOX9 of chondrogenic pellet at day 28. (d) The pellet size of chondrogenic pellets during culture. (e) Deposition of sulfated glycosaminoglycan in chondrogenic pellets during culture. (f) The alteration of chondrogenesis-related gene expression. (g) mRNA expression levels for type 10 collagen in long-term culture and immunostaining for type 10 collagen at day 28 in chondrogenic pellets. (h) Saf-O images of chondrogenic pellet culture with TGFβ3 (10 ng/mL) and various concentrations of BMP2 (0–500 ng/mL) for 28 days. (i) Saf-O images of chondrogenic pellet cultures in TB media with addition of variable serum concentrations for 28 days. Scale bars = 500 μm (a, h, i) and 100 μm (b, c, and high-magnified images of a, h, i). Data are expressed as mean ± SD for three pellet replicates (d–g). Abbreviations: Saf-O: Safranin-O staining; COL2: type 2 collagen; sGAG: sulfated glycosaminoglycan; COL10: type 10 collagen.

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