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

Figure 1

(a) Fabrication of osteogenic iPSC constructs. Mouse GF-iPSCs were cultured under floating conditions in ES medium for 4 days to form EBs. RA was added in the final 2 days. (b) EBs were seeded in wells of a pNIPAAm gel cell chamber (inset, scale bar: 5 mm) and cultured for 2 days in ES medium with RA. (c) Ball-like cell constructs (inset, scale bar: 1 mm) were collected by decreasing the temperature to expand the hydrogel chamber. (d) GF-iPSC constructs were cultured in osteogenic induction medium for 30 days with gentle shaking. (e) Osteogenically induced GF-iPSC construct on day 30 consisting of a white-colored core (asterisk) surrounded by a translucent layer (arrow). Scale bar: 1 mm. (f) von Kossa staining of the osteogenically induced GF-iPSC construct. Scale bar: 1 mm. (g) Live/Dead cell viability assay showed that most cells on the surface of osteogenically induced GF-iPSC constructs were alive with green fluorescence. A few cells were dead with red fluorescence. Scale bar: 100 μm.