Fig. 9
Ptf1a reprograms human foreskin fibroblasts (HFF) into tripotent iNSCs. a, b Ectopic expression of Ptf1a in HFFs by lentiviruses induced the formation of neurospheres. c, d In the absence of doxycycline (Dox), Ptf1a-induced neurosphere cells were capable of forming neurospheres before passage 20 (c), but lost the capacity after passage 20 and became monolayered (d). e–i Ptf1a-induced human neural stem cells (hiNSCs) were highly immunoreactive for PTF1A, SOX2, PAX6, NESTIN, OLIG2, and FABP7. j–p Ptf1a-induced hiNSCs were capable of differentiating into neurons immunoreactive for TUJ1, MAP2, NEUN, TAU, or GABA, astrocytes labeled by GFAP, or oligodendrocytes marked by O1. q Neurons differentiated from hiNSCs were immunoreactive for both Tuj1 and synapsin. Cells in f–q were counterstained with nuclear DAPI. r Voltage-clamp recordings indicated fast activated and inactivated inward sodium currents as well as outward potassium currents on a differentiated neuron. s Current-clamp recordings revealed action potential responses of a differentiated neuron under current injection. t An action potential was induced after depolarization of the neuron. u Spontaneous postsynaptic currents recorded from a differentiated neuron. Scale bars, 80 μm (a–d) and 40 μm (e–q)
