Neural differentiation of HDF-derived iPSCs and characterization of differentiated neurons.

(A) Schematic diagram represents the strategy to differentiate HDF-derived iPSCs into neural lineages using SMAD signaling inhibitors. (B) Morphological changes in HDF-iPSC1 cultures during neural differentiation; undifferentiated HDF-iPSC1 cells on day -1 (i), differentiated cells on day 2 (ii) and day 6 (iii), representative immunofluorescent images show the NEPC on day 6 (iv), NPC on day 9 (v) and post-mitotic neuron on day 28 (vi) of differentiation as examined by PAX6, NESTIN and TUJ1 markers, respectively. Cell nuclei were stained with DAPI. Scale bar = 100 µm. (C) RT-qPCR analysis of pluripotency- and neural-associated genes in NEPCs on day 6 and mature neuron on day 28 of differentiation as compared to those of undifferentiated HDF-iPSC1 cells (arbitrarily set at 1).