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. 2016 Aug 25;7(3):557–570. doi: 10.1016/j.stemcr.2016.07.017

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© 2016 The Authors

This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).

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hiNSCs Rapidly Differentiate into Various Neuronal and Glial Phenotypes

(A) Expression of neuron-specific βIII-tubulin (TUJ1) and glial fibrillary acidic protein (GFAP) in hiNSC clonal lines derived from HFF and hASC at 4 and 14 days. The large panel displays a merged image of TUJ1, GFAP, and DAPI; inlaid insets reveal separate images of TUJ1 (top) and GFAP (bottom). By 4 days, more than 80% of cells stain positive for neuron markers. Scale bars, 100 μm.

(B) hiNSCs spontaneously differentiate into subtypes of neurons. Embryonic rat brain-derived neurons are shown as a positive control. Scale bars, 50 μm.

(C) hiNSCs express post-synaptic proteins at both inhibitory (GEPHYRIN and VGAT) and excitatory (PSD95 and VGLUT1) synapses. Scale bars, 10 μm.

(D) hiNSCs spontaneously express synaptic vesicle protein, SYNAPTOPHYSIN, and voltage-gated sodium channel marker PAN-NAV. Scale bars, 100 μm.

(E) hiNSCs differentiate into multiple types of glia including astrocytes (S100β), oligodendrocytes (O4), and myelin marker positive glia (MBP). Scale bars, 100 μm.

(F) hiNSCs maintain neuronal and glial phenotypes in long-term cultures. Scale bars, 100 μm. Data in (A) and (B) represent mean ± SD of three independent experiments. See also Figures S2–S4 and S6.