Cell nuclei were counterstained with DAPI (blue,a). Approximately 90% of the cells stain positive for the undifferentiated NSC marker nestin and the proliferation marker Ki67. Lack of Oct4 staining indicates that there are no remnant hESCs in the culture (data not shown) (Invitrogen, Manual part no. A11592, MAN0001758). Fluorescence images (20X) of GIBCOR hNSCs that have been cultured in StemProR NSC SFM for three passages, and then allowed to differentiate into neurons, oligodendrocytes, or astrocytes. Upon directed differentiation, cells start to lose the undifferentiated NSC marker, nestin, but stain positive for the differentiated cell type markers Dcx, GalC, and GFAP. Cells were stained for the undifferentiated NSC markers nestin (red, b) and SOX2 (green, c) prior to directed differentiation. Cell were then differentiated into neurons and glial cells, and respectively stained for the neuronal marker Dcx (green, c), for the oligodendrocyte marker GalC (red, d), or for the astrocyte marker, GFAP (green, e). The nuclei were counterstained with DAPI (blue) in panels B–D (Invitrogen, Manual part no. A11592, MAN0001758).