Murine Embryonic Stem Cell-Derived Pyramidal Neurons Integrate into the Cerebral Cortex and Appropriately Project Axons to Subcortical Targets

Abstract

Although embryonic stem (ES) cells have been induced to differentiate into diverse neuronal cell types, the production of cortical projection neurons with the correct morphology and axonal connectivity has not been demonstrated. Here, we show that in vitro patterning is critical for generating neural precursor cells (ES-NPCs) competent to form cortical pyramidal neurons. During the first week of neural induction, these ES-NPCs begin to express genes that are specific for forebrain progenitors; an additional week of differentiation produces mature neurons with many features of cortical pyramidal neurons. After transplantation into the murine cerebral cortex, these specified ES-NPCs manifest the correct dendritic and axonal connectivity for their areal location. ES-NPCs transplanted into the deep layers of the motor cortex differentiate into layer 5 pyramidal neurons and extend axons to distant subcortical targets such as the pons and as far caudal as the pyramidal decussation and descending spinal tract and, importantly, do not extend axons to inappropriate targets such as the superior colliculus (SC). ES-NPCs transplanted into the visual cortex extend axons to the dorsal aspect of the SC and pons but avoid ventral SC and the pyramidal tract, whereas cells transplanted deep into the somatosensory cortex project axons to the ventral SC, avoiding the dorsal SC. Thus, these data establish that ES-derived cortical projection neurons can integrate into anatomically relevant circuits.

WNT-C59, a Small-Molecule WNT Inhibitor, Efficiently Induces Anterior Cortex That Includes Cortical Motor Neurons From Human Pluripotent Stem Cells

“…These PSCs can be unlimitedly expanded and give rise to a wide variety of differentiated cells, making them a promising cell source for treating intractable neurological disorders, including Parkinson's disease [5][6][7] and ALS [8]. In fact, recent studies have shown that mouse [9] or human [10] ESC-derived neurons extend long axons from the motor cortex to the pyramidal tract in neonate mice. However, the efficiency to induce CMNs from ESCs or iPSCs is very low.…”

“…In order to induce cortical neurons from mouse or human ESCs/iPSCs, serum-free embryoid body-like aggregates (SFEB) [11][12][13] or monolayer [9,10,[14][15][16][17] protocols are used. CMNs are located in the frontal/motor area of the anterior cortex, and the inhibition of Wnt signaling is important to induce this area.…”

Telencephalic Neurogenesis Versus Telencephalic Differentiation of Pluripotent Stem Cells

“…By transplantation of dorsally patterned progenitors into postnatal murine cerebral cortex, the production of cortical projection neurons with the correct morphology and axonal connectivity has been demonstrated [98]. The pyramidal neurons express Otx1, Emx1, and Ctip2, corresponding to deep layers neurons.…”

“…The pyramidal neurons express Otx1, Emx1, and Ctip2, corresponding to deep layers neurons. They integrate and appropriately project longdistance axons to subcortical targets, without forming tumors [98].…”

Restoring Neuronal Function After Stroke by Cell Replacement

“…40 Recently, it was demonstrated that ES cell-derived neural populations could generate dorsalized cortical phenotypes by antagonizing sonic hedgehog signaling. 41,42 Significantly, even cortical layer-specific phenotypes were generated, including GABAergic and glutamatergic neurons. After transplantation into neonatal or embryonic mice cortices, these cells showed an area-specific projection into corresponding thalamic nuclei (M1 to ventrolateral thalamic nucleus and S1 to ventrobasal).…”