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dc.contributor.authorSerrano, Felipe
dc.contributor.authorBernard, William
dc.contributor.authorGranata, Alessandra
dc.contributor.authorIyer, Dharini
dc.contributor.authorSteventon, Benjamin
dc.contributor.authorKim, Matthew
dc.contributor.authorVallier, Ludovic
dc.contributor.authorGambardella, Laure
dc.contributor.authorSinha, Sanjay
dc.date.accessioned2018-12-11T00:31:31Z
dc.date.available2018-12-11T00:31:31Z
dc.date.issued2019-01-15
dc.identifier.issn1547-3287
dc.identifier.urihttps://www.repository.cam.ac.uk/handle/1810/286643
dc.description.abstractThe neural crest (NC) is a transient multipotent cell population present during embryonic development. The NC can give rise to multiple cell types and is involved in a number of different diseases. Therefore, the development of new strategies to model NC in vitro enables investigations into the mechanisms involved in NC development and disease. In this study, we report a simple and efficient protocol to differentiate human pluripotent stem cells (HPSC) into NC using a chemically defined media, with basic fibroblast growth factor 2 (FGF2) and the transforming growth factor-β inhibitor SB-431542. The cell population generated expresses a range of NC markers, including P75, TWIST1, SOX10, and TFAP2A. NC purification was achieved in vitro through serial passaging of the population, recreating the developmental stages of NC differentiation. The generated NC cells are highly proliferative, capable of differentiating to their derivatives in vitro and engraft in vivo to NC specific locations. In addition, these cells could be frozen for storage and thawed with no loss of NC properties, nor the ability to generate cellular derivatives. We assessed the potential of the derived NC population to model the neurocristopathy, Treacher Collins Syndrome (TCS), using small interfering RNA (siRNA) knockdown of TCOF1 and by creating different TCOF1+/- HPSC lines through CRISPR/Cas9 technology. The NC cells derived from TCOF1+/- HPSC recapitulate the phenotype of the reported TCS murine model. We also report for the first time an impairment of migration in TCOF1+/- NC and mesenchymal stem cells. In conclusion, the developed protocol permits the generation of the large number of NC cells required for developmental studies, disease modeling, and for drug discovery platforms in vitro.
dc.format.mediumPrint-Electronic
dc.languageeng
dc.publisherMary Ann Liebert Inc
dc.rightsAttribution 4.0 International
dc.rights.urihttps://creativecommons.org/licenses/by/4.0/
dc.subjectPluripotent Stem Cells
dc.subjectChick Embryo
dc.subjectNeural Crest
dc.subjectAnimals
dc.subjectHumans
dc.subjectMandibulofacial Dysostosis
dc.subjectBenzamides
dc.subjectDioxoles
dc.subjectFibroblast Growth Factor 2
dc.subjectAdaptor Proteins, Signal Transducing
dc.subjectNuclear Proteins
dc.subjectPhosphoproteins
dc.subjectTranscription Factors
dc.subjectCell Death
dc.subjectCell Differentiation
dc.subjectCell Movement
dc.subjectTranscription Factor AP-2
dc.subjectSOXE Transcription Factors
dc.subjectNeural Stem Cells
dc.subjectCellular Reprogramming Techniques
dc.subjectTwist-Related Protein 1
dc.titleA Novel Human Pluripotent Stem Cell-Derived Neural Crest Model of Treacher Collins Syndrome Shows Defects in Cell Death and Migration.
dc.typeArticle
prism.endingPage100
prism.issueIdentifier2
prism.publicationDate2019
prism.publicationNameStem Cells Dev
prism.startingPage81
prism.volume28
dc.identifier.doi10.17863/CAM.33955
dcterms.dateAccepted2018-10-29
rioxxterms.versionofrecord10.1089/scd.2017.0234
rioxxterms.versionAM
rioxxterms.licenseref.urihttp://www.rioxx.net/licenses/all-rights-reserved
rioxxterms.licenseref.startdate2019-01
dc.contributor.orcidBernard, William [0000-0002-2622-5115]
dc.contributor.orcidGranata, Alessandra [0000-0001-5051-8201]
dc.contributor.orcidSteventon, Benjamin [0000-0001-7838-839X]
dc.contributor.orcidVallier, Ludovic [0000-0002-3848-2602]
dc.contributor.orcidSinha, Sanjay [0000-0001-5900-1209]
dc.identifier.eissn1557-8534
rioxxterms.typeJournal Article/Review
pubs.funder-project-idWellcome Trust (109408/Z/15/Z)
pubs.funder-project-idMedical Research Council (MC_PC_12009)
pubs.funder-project-idBritish Heart Foundation (PG/17/24/32886)
pubs.funder-project-idWellcome Trust (203151/Z/16/Z)
pubs.funder-project-idBritish Heart Foundation (None)
pubs.funder-project-idBritish Heart Foundation (None)
pubs.funder-project-idNational Centre for the Replacement Refinement and Reduction of Animals in Research (NC/N001540/1)
pubs.funder-project-idBritish Heart Foundation (None)
pubs.funder-project-idBritish Heart Foundation (RG/17/5/32936)
pubs.funder-project-idBritish Heart Foundation (FS/18/46/33663)
pubs.funder-project-idEuropean Commission Horizon 2020 (H2020) Societal Challenges (668294-2 INTENS)
cam.issuedOnline2018-10-30
cam.orpheus.successMon May 31 07:37:15 BST 2021 - The item has an open VoR version.
rioxxterms.freetoread.startdate2100-01-01


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Attribution 4.0 International
Except where otherwise noted, this item's licence is described as Attribution 4.0 International