Journal ArticleDOI
Directed differentiation of human pluripotent cells to neural crest stem cells
Laura Menendez,Michael Kulik,Austin T Page,Sarah S. Park,James D. Lauderdale,Michael L. Cunningham,Michael L. Cunningham,Stephen Dalton +7 more
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TLDR
After completion of this protocol, NCSCs can be used for numerous applications, including the generation of sufficient cell numbers to perform drug screens, for the development of cell therapeutics on an industrial scale and to provide a robust model for human disease.Abstract:
Multipotent neural crest stem cells (NCSCs) have the potential to generate a wide range of cell types including melanocytes; peripheral neurons; and smooth muscle, bone, cartilage and fat cells. This protocol describes in detail how to perform a highly efficient, lineage-specific differentiation of human pluripotent cells to a NCSC fate. The approach uses chemically defined media under feeder-free conditions, and it uses two small-molecule compounds to achieve efficient conversion of human pluripotent cells to NCSCs in ~15 d. After completion of this protocol, NCSCs can be used for numerous applications, including the generation of sufficient cell numbers to perform drug screens, for the development of cell therapeutics on an industrial scale and to provide a robust model for human disease. This protocol can be also be applied to patient-derived induced pluripotent stem cells and thus used to further the knowledge of human disease associated with neural crest development, for example, Treacher-Collins Syndrome.read more
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Journal ArticleDOI
Generation of high-purity human ventral midbrain dopaminergic progenitors for in vitro maturation and intracerebral transplantation
TL;DR: A detailed 16-d protocol for obtaining high-purity ventral midbrain (VM) dopamine (DA) progenitors for intracerebral transplantation into animal models and for in vitro maturation into neurons.
Journal ArticleDOI
Dynamic changes in replication timing and gene expression during lineage specification of human pluripotent stem cells
Juan Carlos Rivera-Mulia,Quinton Buckley,Takayo Sasaki,Jared Zimmerman,Ruth Didier,Kristopher L. Nazor,Jeanne F. Loring,Zheng Lian,Sherman M. Weissman,Allan J. Robins,Thomas C. Schulz,Laura Menendez,Michael Kulik,Stephen Dalton,Haitham Gabr,Tamer Kahveci,David M. Gilbert +16 more
TL;DR: Interestingly, transcriptome data revealed that the well-accepted correlation between early replication and transcriptional activity was restricted to RT-constitutive genes, whereas two-thirds of the genes that switched RT during differentiation were strongly expressed when late replicating in one or more cell types.
Journal Article
A robust method to derive functional neural crest cells from human pluripotent stem cells
Faith R. Kreitzer,Nathan Salomonis,Alice S. Sheehan,Miller Huang,Jason Park,Matthew J. Spindler,Paweena Lizarraga,William A. Weiss,Po-Lin So,Bruce R. Conklin +9 more
TL;DR: A rapid and robust NC differentiation method called "LSB-short" that is based on dual SMAD pathway inhibition that yields high percentages of NC cell populations from multiple human induced pluripotent stem and human embryonic stem cell lines in 8 days.
Journal ArticleDOI
Human pluripotent stem cell-derived brain pericyte-like cells induce blood-brain barrier properties.
Matthew J. Stebbins,Benjamin D. Gastfriend,Scott G. Canfield,Ming-Song Lee,Drew Richards,Madeline G. Faubion,Wan-Ju Li,Richard Daneman,Sean P. Palecek,Eric V. Shusta +9 more
TL;DR: This study generated neural crest stem cells (NCSCs), the embryonic precursor to forebrain pericytes, from hPSCs and subsequently differentiated NCSCs to brain pericyte–like cells, which closely resembled primary human brainpericytes and self-assembled with endothelial cells.
Journal ArticleDOI
MYC Controls Human Pluripotent Stem Cell Fate Decisions through Regulation of Metabolic Flux
Timothy S. Cliff,Tianming Wu,Benjamin R. Boward,Amelia Yin,Hang Yin,John Glushka,James H. Prestegaard,Stephen Dalton +7 more
TL;DR: It is shown that, although metabolic switching occurs during early mesoderm and endoderm differentiation, high glycolytic flux is maintained and, in fact, essential during early ectoderm specification and identifies MYC and MYCN as developmental regulators that couple metabolism to pluripotency and cell fate determination.
References
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Journal ArticleDOI
Highly efficient neural conversion of human ES and iPS cells by dual inhibition of SMAD signaling
Stuart M. Chambers,Christopher A. Fasano,Eirini P. Papapetrou,Mark J. Tomishima,Michel Sadelain,Lorenz Studer +5 more
TL;DR: 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.
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A gene regulatory network orchestrates neural crest formation.
TL;DR: The current understanding of these processes is reviewed and the molecular players that are involved in the neural crest gene regulatory network are discussed.
Journal ArticleDOI
Ectodermal Wnt function as a neural crest inducer.
TL;DR: It is shown that, in avians, Wnt6 is localized in ectoderm and in vivo inhibition of Wnt signaling perturbs neural crest formation, which suggests that Wnt molecules are necessary and sufficient to induce neural crest cells in avian embryos.
Journal ArticleDOI
Multipotentiality of the neural crest.
TL;DR: This work has provided evidence for multipotent stem cells and intermediate precursors in the early NC cell population as well as in various NC derivatives in embryos and even in adult.