In vitro and in vivo characteristics of stem cells from human exfoliated deciduous teeth obtained by enzymatic disaggregation and outgrowth

OBJECTIVE: Stem cells from human exfoliated deciduous teeth (SHED) are a good source of dental tissue for regeneration therapy, and can be obtained using different primary culture methods. The aim of this study was to determine the differences in the in vitro and in vivo characteristics between SHED isolated via enzymatic disaggregation (e-SHED) and outgrowth (o-SHED) primary culture methods. DESIGN: Dental pulp stem cells were isolated from 14 exfoliated deciduous teeth by enzymatic disaggregation (n=7) and outgrowth (n=7). Their proliferation potential and colony-forming ability were evaluated in vitro, as was their mesenchymal stem-cell-marker expression (using flow cytometry), and their differentiation was verified using quantitative real-time PCR (qPCR) and histochemical staining. In addition, the qualitative and quantitative characteristics of the hard tissue that was generated after in vivo transplantation were compared using haematoxylin and eosin staining, immunohistochemical staining, qPCR, and quantitative alkaline phosphatase analysis. RESULTS: The cell-proliferation potential, colony-forming ability, and Stro-1 and CD146 expression were higher in e-SHED than in o-SHED. While the in vitro adipogenic differentiation potential was greater in e-SHED than in o-SHED, the in vitro osteogenic differentiation did not differ significantly between the two cell types. Although in vivo hard tissue formation was greater following transplantation of o-SHED into mice, there was no difference in the quality of hard tissue generated by e-SHED and o-SHED. CONCLUSION: The findings of this study indicate that e-SHED exhibit stronger stemness characteristics, but that o-SHED are more suitable for hard-tissue regeneration therapy in teeth.ope