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. 2003 Dec 17;23(37):11587–11601. doi: 10.1523/JNEUROSCI.23-37-11587.2003

Figure 3.

Figure 3.

EGF promotes the generation and differentiation of cells from embryonic NSCs that antigenically, morphologically, and functionally resemble RGCs. A, Neurospheres grown in EGF and differentiated in basal media (see Materials and Methods) did not give rise to cells resembling RGCs (n = 3) Scale bar, 100 μm. B, Neurospheres grown in EGF and differentiated in EGF-containing media extended long, RGC-like processes (arrows) that appeared to support the migration of immature neurons. Scale bar: B, inset, 20 μm. C, C′, RGC-like processes did not express the neuronal marker TuJ1 (arrow in C, C′; arrowheads indicate immunopositive cells. Scale bar, 50 μm. D, E, RGC-like processes expressed the RGC markers BLBP (D, D′; scale bar, 20 μm) and nestin (E, E′). Arrows indicate RGC process. F, RGC-like processes supported neuronal migration, as shown by time-lapse photography over 195 min (two individual migrating cells are indicated by purple and red arrows; white arrowhead indicates leading process of migrating neuron). Scale bar, 20 μm. G, G′, RGC processes, identified by expression of the RGC marker GLAST (green, arrowhead), supported the migration of cells identified as neurons using the neuron-specific marker TuJ1 (blue, arrow). Scale bar, 20 μm. H, Immature neurons could also be identified on RGC processes by the neuron-specific marker Hu. Scale bar, 20 μm.