FIG. 4.

iPS-dNSCs cultured in definitive media conditions with DLL4 exhibit a more neural-specific gene expression profile compared to iPS-dNSCs cultured in definitive media alone. Selected neural (A), pluripotency (B), and nonectoderm (C) gene markers were examined in the iPS-dNSCs (C5-4A; left and B1-1G; right) cultured with and without DLL4. The addition of DLL4 significantly reduced residual pluripotency and endodermal markers, while maintaining the neural character of the cells. Cyropreserved passage 4 definitive neurospheres for C5-4A cell line cultured in definitive media without DLL4 (upper panels) or with continuous DLL4 treatment (lower panels) were sectioned and stained for NSC markers [(A) nestin, (B) Olig2], a pluripotency marker [(C) Oct4], and a pan-endoderm [(D) Afp] (C5-4A iPSC line). ND denotes not detected; mean±SEM; n=4–5, *P<0.05 compared to iPS-dNSC grown without DLL4; scale bar represents 100 μm.