Adipose tissue-derived stem cell in vitro differentiation in a three-dimensional dental bud structure

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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 trans-differentiate 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. Copyright © 2011 American Society for Investigative Pathology. Published by Elsevier Inc. All rights reserved.

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Ferro, F., Spelat, R., Falini, G., Gallelli, A., D’Aurizio, F., Puppato, E., … Curcio, F. (2011). Adipose tissue-derived stem cell in vitro differentiation in a three-dimensional dental bud structure. American Journal of Pathology, 178(5), 2299–2310. https://doi.org/10.1016/j.ajpath.2011.01.055

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