Generating Functional Sub-cerebral Projection Neurons from Human Induced Pluripotent Stem Cells

In order to define the specification of sub-cerebral projection neurons (SCPNs), we used a transcription activator-like effector nuclease (TALEN)-based approach to target the FEZF2 locus in hIPSCs with an enhanced green fluorescent protein (GFP).

After validating overlap between GFP and FEZF2 expression, we identified gene expression signatures for multipotent neural progenitors and SCPNs. We optimized the differentiation protocol and isolated post-mitotic SCPNs by fluorescence activated cell sorting after three weeks. We then found that GFP-expressing SCPNs exhibited mature electrophysiological properties and glutamatergic synapses after four weeks, which were significantly enhanced by a chemically defined lipid concentrate and co-culture with hIPSC-derived spinal neurons.