Highly efficient neural conversion of human ES and iPS cells by dual inhibition of SMAD signaling

By S. M. Chambers, Christopher A. Fasano, E. Papapetrou, M. Tomishima, Michel Sadelain, L. Studer. Nature Biotechnology, 2009

3,533 citations recorded by the source corpus.

Current neural induction protocols for human embryonic stem (hES) cells rely on embryoid body formation, stromal feeder co-culture or selective survival conditions. Each strategy has considerable drawbacks, such as poorly defined culture conditions, protracted differentiation and low yield. Here we report that the synergistic action of two inhibitors of SMAD signaling, Noggin and SB431542, is sufficient to induce rapid and complete neural conversion of >80% of hES cells under adherent culture conditions. Temporal fate analysis reveals the appearance of a transient FGF5+ epiblast-like stage followed by PAX6+ neural cells competent to form rosettes. Initial cell density determines the ratio of central nervous system and neural crest progeny. Directed differentiation of human induced pluripotent stem (hiPS) cells into midbrain dopamine and spinal motoneurons confirms the robustness and general applicability of the induction protocol. Noggin/SB431542-based neural induction should facilitate the use of hES and hiPS cells in regenerative medicine and disease modeling and obviate the need for protocols based on stromal feeders or embryoid bodies.

Subfields: Biomedical Engineering, Molecular Biology

View Paper: https://www.semanticscholar.org/paper/fc1687f2073117df8cfa5bca60b953288efa33cb

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Citations in the corpus

Epigenomic Analysis of Multi-lineage Differentiation of Human Embryonic Stem Cells (2013, positive): “, 2002), neural progenitor cells (Chambers et al., 2009; Chen et al., 2011), and mesenchymal stem cells (Vodyanik et al.”
Midbrain-like organoids from human pluripotent stem cells contain functional dopaminergic and neuromelanin producing neurons (2016, background): “When neuroectodermal buds were starting to extrude, the hMLOs were embedded in 30 mL of reduced growth factor Matrigel and grown in tissue growth induction medium containing 100 ng/mL SHH-C25II and 100 ng/mL FGF8.”
Stem Cell Organoid Engineering (2016, positive): “, 2013), midbrain dopaminergic neurons (Chambers et al., 2009; Lee et al., 2000; Perrier et al., 2004; Yan et al., 2005), and spinal cord motor neurons (Dimos et al.”
In vitro generation of human pluripotent stem cell derived lung organoids (2015, methodology): “1 is expressed in the lung and thyroid primordium, and PAX8 is expressed in the thyroid primordium, these results suggest that 4 day ActivinA treatment followed by a 4 day NOG/SB treatment biases the cultures towards ventralanterior foregut lineages.”
LRRK2 Mutant iPSC-Derived DA Neurons Demonstrate Increased Susceptibility to Oxidative Stress (2011, positive): “Neuronal and Spontaneous Differentiation Directed differentiation of iPSCs into midbrain dopaminergic neurons was previously described (Chambers et al., 2009) with some adjustments.”
Human pluripotent stem cell-derived neural constructs for predicting neural toxicity (2015, background): “S1) (21), including neural progenitor cells (24), endothelial cells, mesenchymal stem cells (25), and microglia/macrophage precursors ( SI Appendix , SI Materials and Methods ).”
Stoichiometric and temporal requirements of Oct4, Sox2, Klf4, and c-Myc expression for efficient human iPSC induction and differentiation (2009, methodology): “Neural induction was done as previously described (39).”
Human cerebral cortex development from pluripotent stem cells to functional excitatory synapses (2012, methodology): “Results Differentiation to cortical stem cells is retinoid dependent We explored a number of different approaches to direct differentiation of human ES cells to neural stem and progenitor cells of the cerebral cortex in monolayer culture, starting from previously described methods for neura”
Intrinsic Membrane Hyperexcitability of ALS Patient-Derived Motor Neurons (2014, methodology): “All iPSC lines were generated via three-factor (OCT4, SOX2, KLF4) retroviral reprogramming, had a normal karyotype, and differentiated into motor neurons after robust neuralization based on dual SMAD inhibition (Chambers et al., 2009) and specification through exposure to retinoic acid and induction”
Generation of regionally specified neural progenitors and functional neurons from human embryonic stem cells under defined conditions. (2012, methodology): “Recent advances in hESC differentiation and lineage commitment have resulted in improved differentiation protocols for more efficient and synchronized neuralization of hESCs via dual inhibition of SMAD signaling (Chambers et al., 2009; Fasano et al., 2010).”