Scaffold-Free Fabrication of Osteoinductive Cellular Constructs Using Mouse Gingiva-Derived Induced Pluripotent Stem Cells

Figure 3

(a) H&E staining of the osteogenically induced GF-iPSC construct, which consisted of an inner region of unstructured cell mass (inside the white dotted area) and outer osseous-like tissue region (asterisks). Scale bar: 0.5 mm. (b) Magnification of the dotted square in panel (a). Aligned cells were present outside (arrows) and inside (arrow heads) the osseous-like region (asterisk). White circle indicates the inner region of the GF-iPSC construct. Scale bar: 100 μm. (c) Alcian blue staining also indicates the presence of aligned cells outside (arrows) and inside (arrow heads) the osseous-like region (asterisk). White circle indicates the inner region of the GF-iPSC construct. (d)–(h) Staining for HIF-1α ((e) green fluorescence), type II collagen ((f) green fluorescence), osteocalcin ((h) red fluorescence), and nuclei ((d)–(h) blue fluorescence). Staining with nonspecific control IgGs as primary antibodies was used as a negative control (d and g). Aligned cells (arrows), osseous region (asterisks), and inner region (circles) of the cellular construct are indicated. Scale bars: 100 μm.

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