Fig. 4

Spontaneous and directed GMP-compatible differentiation of lt-NES. a Spontaneous differentiation of NES lines into neurons under GMP-compatible conditions. Spontaneous neurogenic potential shown by phase contrast (scale bars 100 μm) and immunostaining for neuronal marker BIII Tubulin (NES7; scale bar 400 μm). Typical hindbrain bias of lt-NES shown by expression of GABAergic markers GABA (scale bars 400 μm). b Directed differentiation of NES lines into midbrain dopaminergic neurons under GMP-compatible conditions. Immunostaining images showing positivity for dopaminergic markers tyrosine hydroxylase (TH) and Nurr1. Scale bars 400 μm (200 μm for AF22 Nurr1). c Directed differentiation of NES lines into motoneurons under GMP-compatible conditions showing motor neuron marker HB9 expression by immunofluorescence. Scale bars 100 μm