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Physiological normoxia and absence of EGF is required for the long-term propagation of anterior neural precursors from human pluripotent cells.

TitlePhysiological normoxia and absence of EGF is required for the long-term propagation of anterior neural precursors from human pluripotent cells.
Publication TypeJournal Article
Year of Publication2014
AuthorsBilican B, Livesey MR, Haghi G, Qiu J, Burr K, Siller R, Hardingham GE, Wyllie DJA, Chandran S
JournalPLoS One
Volume9
Issue1
Paginatione85932
Date Published2014
ISSN1932-6203
KeywordsCell Differentiation, Cells, Cultured, Cerebral Cortex, Epidermal Growth Factor, Fibroblast Growth Factor 2, Humans, Neural Stem Cells, Neurons, Oxygen, Pluripotent Stem Cells, Signal Transduction, Synapses, Time Factors
Abstract

Widespread use of human pluripotent stem cells (hPSCs) to study neuronal physiology and function is hindered by the ongoing need for specialist expertise in converting hPSCs to neural precursor cells (NPCs). Here, we describe a new methodology to generate cryo-preservable hPSC-derived NPCs that retain an anterior identity and are propagatable long-term prior to terminal differentiation, thus abrogating regular de novo neuralization. Key to achieving passagable NPCs without loss of identity is the combination of both absence of EGF and propagation in physiological levels (3%) of O2. NPCs generated in this way display a stable long-term anterior forebrain identity and importantly retain developmental competence to patterning signals. Moreover, compared to NPCs maintained at ambient O2 (21%), they exhibit enhanced uniformity and speed of functional maturation, yielding both deep and upper layer cortical excitatory neurons. These neurons display multiple attributes including the capability to form functional synapses and undergo activity-dependent gene regulation. The platform described achieves long-term maintenance of anterior neural precursors that can give rise to forebrain neurones in abundance, enabling standardised functional studies of neural stem cell maintenance, lineage choice and neuronal functional maturation for neurodevelopmental research and disease-modelling.

DOI10.1371/journal.pone.0085932
Alternate JournalPLoS ONE
PubMed ID24465796
PubMed Central IDPMC3895023
Grant List092742/Z/10/Z / / Wellcome Trust / United Kingdom
/ / Medical Research Council / United Kingdom
Publication institute
CRM