Fig 1: Differentiation of nucleostemin-expressed cells (early P2 day 14) to all three germ layers proved their stemness. Morphological analysis (magnification, ×10; scale bar, 100 µm) validated the cell differentiation, immunocytochemical analysis (magnification, ×20; scale bar, 50 µm) indicated the cell specificity, and semi-quantitative gene analysis validated their functionality. (A) Morphological analysis of neuroectoderm neuronal lineage cells, (B) immunocytochemical analysis of neuronal marker MAP2 at day 18 after neural induction, and (C) neural-specific gene expression. (D) Morphological analysis of definitive endoderm hepatocytes, (E) immunocytochemical analysis of their marker Sox17 at day 28 after induction, and (F) hepatocyte-specific gene expression. (G) Morphological analysis of mesoderm osteogenic cells, (H) immunocytochemical analysis of the marker collagen I at day 18 after induction, and (I) osteogenic gene expression. S1 and S2 represent samples 1 and 2, respectively. MAP2, microtubule-associated protein 2; Sox17, SRY-box 17.