title: In vitro and in vivo characteristics of stem cells from human exfoliated deciduous teeth obtained by enzymatic disaggregation and outgrowth.
abstract: 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.
  
  
  METHODS
  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.
  
  
  CONCLUSIONS
  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.
authors: Department of Pediatric Dentistry, Oral Science Research Center, College of Dentistry, Yonsei University, Seoul, Republic of Korea.
authors: Mijeong
authors: M
authors: Jeon
authors: Department of Pediatric Dentistry, Oral Science Research Center, College of Dentistry, Yonsei University, Seoul, Republic of Korea.
authors: Je Seon
authors: JS
authors: Song
authors: Department of Pediatric Dentistry, Oral Science Research Center, College of Dentistry, Yonsei University, Seoul, Republic of Korea.
authors: Byung-Jai
authors: BJ
authors: Choi
authors: Department of Pediatric Dentistry, Oral Science Research Center, College of Dentistry, Yonsei University, Seoul, Republic of Korea.
authors: Hyung-Jun
authors: HJ
authors: Choi
authors: Department of Oral Biology, College of Dentistry, Yonsei University, Seoul, Republic of Korea.
authors: Dong-Min
authors: DM
authors: Shin
authors: Division in Anatomy & Developmental Biology, Department of Oral Biology, College of Dentistry, Yonsei University, Seoul, Republic of Korea.
authors: Han-Sung
authors: HS
authors: Jung
authors: Department of Pediatric Dentistry, Oral Science Research Center, College of Dentistry, Yonsei University, Seoul, Republic of Korea. Electronic address: ksodds@yuhs.ac.
authors: Seong-Oh
authors: SO
authors: Kim
mesh_terms: D000469:Alkaline Phosphatase / Q000032:analysis; D000818:Animals; D002454:Cell Differentiation / Q000502:physiology; D049109:Cell Proliferation / Q000502:physiology; D002648:Child; D002675:Child, Preschool; D005260:Female; D005434:Flow Cytometry; D006801:Humans; D066298:In Vitro Techniques; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D010012:Osteogenesis / Q000502:physiology; D060888:Real-Time Polymerase Chain Reaction; D012038:Regeneration / Q000502:physiology; D044968:Regenerative Medicine; D013194:Staining and Labeling; D033581:Stem Cell Transplantation; D013234:Stem Cells / Q000166:cytology* / Q000201:enzymology; D014080:Tooth Exfoliation; D014094:Tooth, Deciduous / Q000166:cytology*
chemical_list: D000469:Alkaline Phosphatase
keywords: Enzymatic disaggregation; Hard tissues; In vivo transplantation; Outgrowth; Stem cells from human exfoliated deciduous teeth (SHED)