title: ES cells-derived ectomesenchymal cells for tooth engineering.
abstract: Recent progresses in stem cell biology and tissue engineering allow considering the possible development of new therapies for compensating the dental tissue losses associated with traumas, pathologies or ageing. The possibility of generating a tooth by mimicking development through reassociations between dental epithelial cells and ectomesenchymal cells derived from the neural crest (NC) has been demonstrated in the mouse. In the search of cell sources to be used for a human transfer, pluripotent stem cells could represent a good alternative. Our study thus focuses on obtaining, ectomesenchymal cells from pluripotent ES cells, capable of promoting tooth histomorphogenesis, when reassociated with a competent dental epithelium. To this end, two ES differentiation protocols, using cyclopamine or a combination of FGF2 and BMP4, have been developed and tested for their capacity to generate such cells. The differentiated ES cells were characterized by quantitative RT-PCR. Both protocols led the cells to acquire in 10 days a mesenchymal-like cell morphology. Rapidly after induction, the cells loose their expression of pluripotent genes while sequentially activating typical NC specifiers. However, the kinetics of gene activation differed between the 2 protocols. Interestingly, Twist, a gene whose expression in the NC is associated with a commitment towards an ectomesenchymal fate, is only activated under the influence of FGF2 and BMP4. Reassociation experiments with a competent epithelium will allow testing the odontogenic potential of the differentiated ES cells. These experiments performed in the mouse system should allow defining a strategy for obtaining odontogenic competent human cells.
authors.affiliation: EA 2496, Faculté de Chirurgie Dentaire, Université Paris Descartes, Montrouge, France.
authors.forename: S
authors.initials: S
authors.lastname: Acuña-Mendoza
authors.affiliation: CNRS UMR 7241/INSERM U1050, Collège de France, Paris, France.
authors.forename: S
authors.initials: S
authors.lastname: Martin
authors.affiliation: EA 2496, Faculté de Chirurgie Dentaire, Université Paris Descartes, Montrouge, France.
authors.forename: S
authors.initials: S
authors.lastname: Ribes
authors.affiliation: Faculté de Chirurgie Dentaire, Université de Strasbourg, Strasbourg, France.
authors.forename: L
authors.initials: L
authors.lastname: Keller
authors.affiliation: EA 2496, Faculté de Chirurgie Dentaire, Université Paris Descartes, Montrouge, France.
authors.forename: C
authors.initials: C
authors.lastname: Chaussain
authors.affiliation: CNRS UMR 7241/INSERM U1050, Collège de France, Paris, France.
authors.forename: L
authors.initials: L
authors.lastname: Lebrin
authors.affiliation: Faculté de Chirurgie Dentaire, Université de Strasbourg, Strasbourg, France.
authors.forename: H
authors.initials: H
authors.lastname: Lesot
authors.affiliation: EA 2496, Faculté de Chirurgie Dentaire, Université Paris Descartes, Montrouge, France.
authors.forename: A
authors.initials: A
authors.lastname: Poliard
mesh_terms: D000818:Animals; D055415:Bone Morphogenetic Protein 4 / Q000494:pharmacology; D018929:Cell Culture Techniques; D002454:Cell Differentiation / Q000187:drug effects; D002465:Cell Movement / Q000502:physiology; D048430:Cell Shape; D002478:Cells, Cultured; D003470:Culture Media; D004475:Ectoderm / Q000166:cytology* / Q000187:drug effects; D016222:Fibroblast Growth Factor 2 / Q000494:pharmacology; D005455:Fluorescent Antibody Technique; D008648:Mesoderm / Q000166:cytology* / Q000187:drug effects; D051379:Mice; D009432:Neural Crest / Q000166:cytology; D009805:Odontogenesis / Q000502:physiology*; D010641:Phenotype; D039904:Pluripotent Stem Cells / Q000166:cytology* / Q000187:drug effects; D060888:Real-Time Polymerase Chain Reaction; D020133:Reverse Transcriptase Polymerase Chain Reaction; D023822:Tissue Engineering / Q000379:methods*; D015533:Transcriptional Activation / Q000235:genetics; D051879:Twist-Related Protein 1 / Q000187:drug effects / Q000235:genetics; D014704:Veratrum Alkaloids / Q000494:pharmacology
chemical_list: D055415:Bone Morphogenetic Protein 4; D003470:Culture Media; D051879:Twist-Related Protein 1; D014704:Veratrum Alkaloids; D016222:Fibroblast Growth Factor 2; C000541:cyclopamine