Adipose tissue-derived stem cell in vitro differentiation in a three-dimensional dental bud structure.
Ferro, Federico ; Spelat, Renza ; Falini, Giuseppe ; et al. ; - ASI Sponsor
May - 2011
ISSN : 1525-2191 ;
journal : The American journal of pathology

Issue : 5
type: Article Journal

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.

keywords : Adipose Tissue,Adipose Tissue: cytology,Adult,Blotting,Cell Lineage,Cell Separation,Cell Transdifferentiation,Cell Transdifferentiation: physiology,Electron,Flow Cytometry,Fluorescent Antibody Technique,Humans,Immunohistochemistry,Mesenchymal Stem Cells,Mesenchymal Stem Cells: cytology,Microscopy,Odontogenesis,Odontogenesis: physiology,Reverse Transcriptase Polymerase Chain Reaction,Tooth,Tooth: cytology,Tooth: growth \& development,Transmission,Western,X-Ray Diffraction