title: Early acquisition of neural crest competence during hESCs neuralization.
abstract: BACKGROUND
  Neural crest stem cells (NCSCs) are a transient multipotent embryonic cell population that represents a defining characteristic of vertebrates. The neural crest (NC) gives rise to many derivatives including the neurons and glia of the sensory and autonomic ganglia of the peripheral nervous system, enteric neurons and glia, melanocytes, and the cartilaginous, bony and connective tissue of the craniofacial skeleton, cephalic neuroendocrine organs, and some heart vessels.
  
  
  RESULTS
  We present evidence that neural crest (NC) competence can be acquired very early when human embryonic stem cells (hESCs) are selectively neuralized towards dorsal neuroepithelium in the absence of feeder cells in fully defined conditions. When hESC-derived neurospheres are plated on fibronectin, some cells emigrate onto the substrate. These early migratory Neural Crest Stem Cells (emNCSCs) uniformly upregulate Sox10 and vimentin, downregulate N-cadherin, and remodel F-actin, consistent with a transition from neuroepithelium to a mesenchymal NC cell. Over 13% of emNCSCs upregulate CD73, a marker of mesenchymal lineage characteristic of cephalic NC and connexin 43, found on early migratory NC cells. We demonstrated that emNCSCs give rise in vitro to all NC lineages, are multipotent on clonal level, and appropriately respond to developmental factors. We suggest that human emNCSC resemble cephalic NC described in model organisms. Ex vivo emNCSCs can differentiate into neurons in Ret.k(-) mouse embryonic gut tissue cultures and transplanted emNCSCs incorporate into NC-derived structures but not CNS tissues in chick embryos.
  
  
  CONCLUSIONS
  These findings will provide a framework for further studying early human NC development including the epithelial to mesenchymal transition during NC delamination.
authors.affiliation: Sanford-Burnham Institute for Medical Research, La Jolla, California, United States of America.
authors.forename: Carol Lynn
authors.initials: CL
authors.lastname: Curchoe
authors.forename: Jochen
authors.initials: J
authors.lastname: Maurer
authors.forename: Sonja J
authors.initials: SJ
authors.lastname: McKeown
authors.forename: Giulio
authors.initials: G
authors.lastname: Cattarossi
authors.forename: Flavio
authors.initials: F
authors.lastname: Cimadamore
authors.forename: Mats
authors.initials: M
authors.lastname: Nilbratt
authors.forename: Evan Y
authors.initials: EY
authors.lastname: Snyder
authors.forename: Marianne
authors.initials: M
authors.lastname: Bronner-Fraser
authors.forename: Alexey V
authors.initials: AV
authors.lastname: Terskikh
mesh_terms: D000818:Animals; D002454:Cell Differentiation / Q000235:genetics; D019070:Cell Lineage; D002465:Cell Movement / Q000235:genetics; D002478:Cells, Cultured; D002642:Chick Embryo; D016000:Cluster Analysis; D053595:Embryonic Stem Cells / Q000166:cytology / Q000378:metabolism*; D058750:Epithelial-Mesenchymal Transition / Q000235:genetics; D020869:Gene Expression Profiling; D006801:Humans; D007150:Immunohistochemistry; D007413:Intestinal Mucosa / Q000378:metabolism; D007422:Intestines / Q000196:embryology; D051379:Mice; D039902:Multipotent Stem Cells / Q000166:cytology / Q000378:metabolism*; D009432:Neural Crest / Q000166:cytology / Q000378:metabolism*; D058953:Neural Stem Cells / Q000166:cytology / Q000378:metabolism*; D020411:Oligonucleotide Array Sequence Analysis; D009924:Organ Culture Techniques; D000072041:PAX3 Transcription Factor; D051761:Paired Box Transcription Factors / Q000235:genetics / Q000378:metabolism; D020800:Receptor, Nerve Growth Factor / Q000235:genetics / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D055757:SOXE Transcription Factors / Q000235:genetics / Q000378:metabolism
chemical_list: D000072041:PAX3 Transcription Factor; C489409:PAX3 protein, human; D051761:Paired Box Transcription Factors; D020800:Receptor, Nerve Growth Factor; C527923:SOX10 protein, human; D055757:SOXE Transcription Factors