title: Adipose tissue-derived stem cell in vitro differentiation in a three-dimensional dental bud structure.
abstract: Tooth morphogenesis requires sequential and reciprocal interactions between the cranial neural crest-derived mesenchymal cells and the stomadial epithelium, which regulate tooth morphogenesis and differentiation. We show how mesenchyme-derived single stem cell populations can be induced to transdifferentiate in vitro in a structure similar to a dental bud. The presence of stem cells in the adipose tissue has been previously reported. We incubated primary cultures of human adipose tissue-derived stem cells in a dental-inducing medium and cultured the aggregates in three-dimensional conditions. Four weeks later, cells formed a three-dimensional organized structure similar to a dental bud. Expression of dental tissue-related markers was tested assaying lineage-specific mRNA and proteins by RT-PCR, immunoblot, IHC, and physical-chemical analysis. In the induction medium, cells were positive for ameloblastic and odontoblastic markers as both mRNAs and proteins. Also, cells expressed epithelial, mesenchymal, and basement membrane markers with a positional relationship similar to the physiologic dental morphogenesis. Physical-chemical analysis revealed 200-nm and 50-nm oriented hydroxyapatite crystals as displayed in vivo by enamel and dentin, respectively. In conclusion, we show that adipose tissue-derived stem cells in vitro can transdifferentiate to produce a specific three-dimensional organization and phenotype resembling a dental bud even in the absence of structural matrix or scaffold to guide the developmental process.
authors.affiliation: Department of Pathology and Experimental and Clinical Medicine, University of Udine, Udine, Italy.
authors.forename: Federico
authors.initials: F
authors.lastname: Ferro
authors.forename: Renza
authors.initials: R
authors.lastname: Spelat
authors.forename: Giuseppe
authors.initials: G
authors.lastname: Falini
authors.forename: Annarita
authors.initials: A
authors.lastname: Gallelli
authors.forename: Federica
authors.initials: F
authors.lastname: D'Aurizio
authors.forename: Elisa
authors.initials: E
authors.lastname: Puppato
authors.forename: Maura
authors.initials: M
authors.lastname: Pandolfi
authors.forename: Antonio Paolo
authors.initials: AP
authors.lastname: Beltrami
authors.forename: Daniela
authors.initials: D
authors.lastname: Cesselli
authors.forename: Carlo Alberto
authors.initials: CA
authors.lastname: Beltrami
authors.forename: Francesco Saverio
authors.initials: FS
authors.lastname: Ambesi-Impiombato
authors.forename: Francesco
authors.initials: F
authors.lastname: Curcio
mesh_terms: D000273:Adipose Tissue / Q000166:cytology*; D000328:Adult; D015153:Blotting, Western; D019070:Cell Lineage; D002469:Cell Separation; D054338:Cell Transdifferentiation / Q000502:physiology*; D005434:Flow Cytometry; D005455:Fluorescent Antibody Technique; D006801:Humans; D007150:Immunohistochemistry; D066298:In Vitro Techniques; D059630:Mesenchymal Stem Cells / Q000166:cytology*; D046529:Microscopy, Electron, Transmission; D009805:Odontogenesis / Q000502:physiology*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014070:Tooth / Q000166:cytology* / Q000254:growth & development; D014961:X-Ray Diffraction