ArticleNature protocols2011
Efficient derivation of NPCs, spinal motor neurons and midbrain dopaminergic neurons from hESCs at 3% oxygen.
Article in Nature protocols, 2011. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 26 papers.
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Who cites it
26 citing papers in PubMed.
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- Acoustofluidic bioassembly induced morphogenesis for therapeutic tissue fabrication.Nature communications · 2025Article
- 3D human induced pluripotent stem cell-derived bioengineered skeletal muscles for tissue, disease and therapy modeling.Nature protocols · 2023Review
- Human muscle production in vitro from pluripotent stem cells: Basic and clinical applications.Seminars in cell & developmental biology · 2021Review
- Disease-specific phenotypes in iPSC-derived neural stem cells with POLG mutations.EMBO molecular medicine · 2020Article
- Nup133 and ERα mediate the differential effects of hyperoxia-induced damage in male and female OPCs.Molecular and cellular pediatrics · 2020Article
- Modeling neuronopathic storage diseases with patient-derived culture systems.Neurobiology of disease · 2019Review
- Human Embryonic Stem Cell-Derived Neural Lineages asBioMed research international · 2019Article
- Review
- Three-Dimensional Human iPSC-Derived Artificial Skeletal Muscles Model Muscular Dystrophies and Enable Multilineage Tissue Engineering.Cell reports · 2018Article
- Hypoxic Culture Promotes Dopaminergic-Neuronal Differentiation of Nasal Olfactory Mucosa Mesenchymal Stem Cells via Upregulation of Hypoxia-Inducible Factor-1α.Cell transplantation · 2017Article
- Hypoxia Epigenetically Confers Astrocytic Differentiation Potential on Human Pluripotent Cell-Derived Neural Precursor Cells.Stem cell reports · 2017Article
- Evidence for evolutionary divergence of activity-dependent gene expression in developing neurons.eLife · 2016Article
- Pluripotent Human embryonic stem cell derived neural lineages for in vitro modelling of enterovirus 71 infection and therapy.Virology journal · 2016Article
- Establishment of Human Neural Progenitor Cells from Human Induced Pluripotent Stem Cells with Diverse Tissue Origins.Stem cells international · 2016Article
- Cell cycle and p53 gate the direct conversion of human fibroblasts to dopaminergic neurons.Nature communications · 2015Article
- Differentiation of Human Neural Stem Cells into Motor Neurons Stimulates Mitochondrial Biogenesis and Decreases Glycolytic Flux.Stem cells and development · 2015Article
- Accelerated neuronal differentiation toward motor neuron lineage from human embryonic stem cell line (H9).Tissue engineering. Part C, Methods · 2015Article
- Influence of oxygen tension on dopaminergic differentiation of human fetal stem cells of midbrain and forebrain origin.PloS one · 2014Article
- Article
Corrections and comments
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Authors and funding
9 authors.
Funding
Abstract
This protocol has been designed to generate neural precursor cells (NPCs) from human embryonic stem cells (hESCs) using a physiological oxygen (O(2)) level of 3% (previously termed hypoxia) and chemically defined conditions. The first stage involves suspension culture of hESC colonies at 3% O(2), where they acquire a neuroepithelial identity over a period of 2 weeks. This timescale is comparable to that observed at 20% O(2), but survival is enhanced. Sequential application of retinoic acid and purmorphamine (PM), from day 14 to day 28, directs differentiation toward spinal motor neurons. Alternatively, addition of fibroblast growth factor-8 and PM generates midbrain dopaminergic neurons. OLIG2 (encoding oligodendrocyte lineage transcription factor 2) induction in motor neuron precursors is twofold greater than that at 20% O(2), whereas EN1 (encoding engrailed homeobox 1) expression is enhanced fivefold. NPCs (at 3% O(2)) can be differentiated into all three neural lineages, and such cultures can be maintained long term in the absence of neurotrophins. The ability to generate defined cell types at 3% O(2) should represent a significant advancement for in vitro disease modeling and potentially for cell-based therapies.
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Registered trials
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