Figure 2.
Figure 2. Maintenance of pluripotency and differentiation capability of NANOG-overexpressing hESC sublines. (A) Assessment of pluripotency in NANOG-overexpressing and control sublines as well as wild-type cells by AP staining assay. The data represent the mean ± SEM (error bars) from three independent experiments. (B) Histological analysis of teratomas formed from grafted colonies of hESCs (hES-NCL1 NANOG) in SCID mice. (a) Low-power light micrograph showing heterogeneous structure within the body of the teratoma and the presence of a diverse range of different tissue types. (b) Cartilage (cart) and bone. (c) Kidney glomeruli (g) and associated tubules (t). (d) Large neural ganglion (ng) with connecting nerve fibers (nf) shown by a black arrow. (e) Transverse section through primitive intestine (in) with accompanying submucosal muscle layer (m). (f) Villous-type structure (v) lined with mucus-secreting cells resembling an intestinal epithelium (ep). Histological staining: Weigert's (a and b) and hematoxylin and eosin (c–f). Bars: (a) 400 μm; (b and e) 150 μm; (c, d, and f) 75 μm. (C) Histological analysis of teratomas formed from grafted colonies of hESCs (hES-NCL1 control) in SCID mice. (a) Low-power light micrograph showing heterogeneous structure within the body of the teratoma and the presence of a diverse range of different tissue types. (b) Cartilage. (c) Bone. (d) Small neural ganglia were clearly identifiable between layers of connective tissue (cn). (e) Kidney glomeruli and associated tubules. (f) Higher magnification image of a secretory cell resembling an intestinal epithelium producing mucus from goblet cells (gc) marked by two black arrows. Histological staining: Weigert's (a–c) and hematoxylin and eosin (d–f). Bars: (a) 400 μm; (b–e) 75 μm; (f) 25 μm. (D) Immunohistochemical analysis of teratomas formed from grafted colonies of hES–NCL1 NANOG in SCID mice. hES–NCL1 NANOG cells were found to be pluripotent, and the teratomas formed were composed of cell types from all three germ layers. (a) AFP (endoderm). (b) Nestin-positive cells (ectoderm). (c) SMA-positive cells (mesoderm). (d) Negative control. Bars, 150 μm.

Maintenance of pluripotency and differentiation capability of NANOG-overexpressing hESC sublines. (A) Assessment of pluripotency in NANOG-overexpressing and control sublines as well as wild-type cells by AP staining assay. The data represent the mean ± SEM (error bars) from three independent experiments. (B) Histological analysis of teratomas formed from grafted colonies of hESCs (hES-NCL1 NANOG) in SCID mice. (a) Low-power light micrograph showing heterogeneous structure within the body of the teratoma and the presence of a diverse range of different tissue types. (b) Cartilage (cart) and bone. (c) Kidney glomeruli (g) and associated tubules (t). (d) Large neural ganglion (ng) with connecting nerve fibers (nf) shown by a black arrow. (e) Transverse section through primitive intestine (in) with accompanying submucosal muscle layer (m). (f) Villous-type structure (v) lined with mucus-secreting cells resembling an intestinal epithelium (ep). Histological staining: Weigert's (a and b) and hematoxylin and eosin (c–f). Bars: (a) 400 μm; (b and e) 150 μm; (c, d, and f) 75 μm. (C) Histological analysis of teratomas formed from grafted colonies of hESCs (hES-NCL1 control) in SCID mice. (a) Low-power light micrograph showing heterogeneous structure within the body of the teratoma and the presence of a diverse range of different tissue types. (b) Cartilage. (c) Bone. (d) Small neural ganglia were clearly identifiable between layers of connective tissue (cn). (e) Kidney glomeruli and associated tubules. (f) Higher magnification image of a secretory cell resembling an intestinal epithelium producing mucus from goblet cells (gc) marked by two black arrows. Histological staining: Weigert's (a–c) and hematoxylin and eosin (d–f). Bars: (a) 400 μm; (b–e) 75 μm; (f) 25 μm. (D) Immunohistochemical analysis of teratomas formed from grafted colonies of hES–NCL1 NANOG in SCID mice. hES–NCL1 NANOG cells were found to be pluripotent, and the teratomas formed were composed of cell types from all three germ layers. (a) AFP (endoderm). (b) Nestin-positive cells (ectoderm). (c) SMA-positive cells (mesoderm). (d) Negative control. Bars, 150 μm.

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