Generation of Regionally Specified Neural Progenitors and Functional Neurons from Human Embryonic Stem Cells under Defined Conditions

摘要

SummaryTo model human neural-cell-fate specification and to provide cells for regenerative therapies, we have developed a method to generate human neural progenitors and neurons from human embryonic stem cells, which recapitulates human fetal brain development. Through the addition of a small molecule that activates canonical WNT signaling, we induced rapid and efficient dose-dependent specification of regionally defined neural progenitors ranging from telencephalic forebrain to posterior hindbrain fates. Ten days after initiation of differentiation, the progenitors could be transplanted to the adult rat striatum, where they formed neuron-rich and tumor-free grafts with maintained regional specification. Cells patterned toward a ventral midbrain (VM) identity generated a high proportion of authentic dopaminergic neurons after transplantation. The dopamine neurons showed morphology, projection pattern, and protein expression identical to that of human fetal VM cells grafted in parallel. VM-patterned but not forebrain-patterned neurons released dopamine and reversed motor deficits in an animal model of Parkinson's disease.Graphical AbstractFigure optionsView in workspaceDownload full-size imageDownload as PowerPoint slideHighlights? A rapid and defined protocol for region-specific neuralization of hESCs is presented ? Caudalization is obtained by graded chemical activation of WNT signaling ? Neuron-rich and tumor-free grafts are obtained after only 10 days of predifferentiation ? VM-specific grafts reverse motor deficits and release dopamine in an animal model of Parkinson's disease.