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. 2016 Sep 7;6(2):458–470. doi: 10.5966/sctm.2016-0177

Figure 2.

Figure 2

Conditional induction of GMSCs into iNPCs. GMSCs were cultured in neurobasal medium supplemented with 1% N‐2 Supplement, 2% B27, 20 ng/ml EGF, and 20 ng/ml bFGF under suspension culture conditions for 3 days. (A): Spheroid cells were dissociated and cultured in polyornithine/laminin‐coated 6‐well plates with the same medium for 7 days. Morphological changes were observed under a microscope. (B): Subculture of GMSC‐derived spheroid and iNPCs. Scale bars = 100 µm (×10) (C,D): Immunofluorescence studies on the expression of Nestin, SOX1, PAX6, and Vimentin in GMSC‐derived iNPC colonies. Nuclei were counterstained with DAPI (blue). Scale bars = 20 µm (×40). (E): Percentage of cells positive for Nestin, SOX1, PAX6, and Vimentin in GMSC‐derived iNPCs determined by flow cytometry. (F) Western blot analysis of the expression of Nestin, SOX1, PAX6, and Vimentin GMSC‐derived iNPCs; β‐actin was used as the internal control. Abbreviations: bFGF, basic fibroblast growth factor; DAPI, 4′,6‐diamidino‐2‐phenylindole; 3D, three‐dimensional; EGF, epidermal growth factor; GMSCs, gingiva‐derived mesenchymal stem cells; iNPCs, induced neural progenitor‐like cells.