Chemically defined differentiation of human pluripotent stem cells to hindbrain and spinal cord neural stem cells with defined regional identities
Year: 2015
Type: article
Abstract: This chemically de ned protocol facilitates the differentiation of human pluripotent stem cells \(hPSCs) to highly pure neural stem cells \(NSCs) characteristic of diverse anatomical regions throughout the posterior central nervous system \(CNS).Whereas previous protocols generated NSC cultures of predominantly cervical spinal cord or heterogeneous regional identity, this protocol can generate NSCs with highly uniform \(60-90% purity), distinct positional identity from discrete regions spanning the rostrocaudal neuraxis of the hindbrain thru cervical, thoracic, and lumbosacral spinal cord.The process relies on inducing colinear _HOX_ activation in a pre-NSC culture by modulating Wnt/β-catenin, broblast growth factor \(FGF), and growth differentiation factor \(GDF) signaling.Then, the application of retinoic acid \(RA) halts _HOX_ activation and transitions the cultures to PAX6+ NSCs expressing HOX factors characteristic of speci c hindbrain and spinal cord locations.Region-speci c NSCs, which may have clinical utility, can be generated within 6-14 days depending on their rostrocaudal location.
Source: Protocol Exchange
Institution: University of Wisconsin–Madison
Language: English
Subfield: Molecular Biology
Domain: Life Sciences
Open Access status: hybrid
OpenAlex ID: W2891561027