Chunk #24 — Results — Induction of neural stem cells (NSCs) from iPSCs and generation of human 2D neuronal culture and 3D neuro-spheroids (3DSs) from NSCs
staining clearly indicated the withdrawal of the neurons from the cell cycle and the initiation of terminal differentiation of the neurons [30]. Almost all of neuronal stem cells were differentiated into neurons, as class III β-tubulin is a microtubule element expressed exclusively in neurons [31]. Our spheroids were also immunoreactive with GFAP antibody that we have used to stain human brain tissue in our previous studies [32], suggesting a mixed population of neurons and astrocytes/glia cells. MAP2 is an abundant neuronal cytoskeletal protein that binds to tubulin and associates with and stabilizes microtubules [33], and our differentiated neurons exhibited similar MAP2 staining. Expression of transcription factor PAX6, an early marker of neuronal differentiation [34], drove the differentiation of all five stem cell lines to neurons.
