Journal of the korean academy of Pediatric Dentistry (대한소아치과학회지)

Korean Academy of Pediatric Dentistry (대한소아치과학회)
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Characterization of Human Dental Pulp Cells from Supernumerary Teeth by Using Flow Cytometry Analysis

유세포 분석을 통한 과잉치 치수 유래 세포의 줄기세포 특성 연구

  • You, Yonsook (Misa Seoul S Dental Clinic) ;
  • Kim, Jongbin (Department of Pediatric Dentistry, School of Dentistry, Dankook University) ;
  • Shin, Jisun (Department of Pediatric Dentistry, School of Dentistry, Dankook University) ;
  • Lee, June-Haeng (Department of Pediatric Dentistry, School of Dentistry, Dankook University) ;
  • Kim, Jongsoo (Department of Pediatric Dentistry, School of Dentistry, Dankook University)
  • 류연숙 (미사서울치과) ;
  • 김종빈 (단국대학교 치과대학 소아치과학교실) ;
  • 신지선 (단국대학교 치과대학 소아치과학교실) ;
  • 이준행 (단국대학교 치과대학 소아치과학교실) ;
  • 김종수 (단국대학교 치과대학 소아치과학교실)
  • Received : 2019.03.27
  • Accepted : 2019.06.25
  • Published : 2019.08.31
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Abstract

The aim of this study was to analyze cells from human dental pulp tissue of impacted supernumerary teeth as stem cells with flow cytometry. Human dental pulp cells from 15 supernumerary teeth were identified their characteristics as stem cells by expression of mesenchymal stem cell markers through flow cytometry analysis at passage 3 and passage 10. Cluster of differentiation (CD) 73, CD 90, CD 34, CD 45 and STRO-1 cell surface markers were used to figure out characteristics of dental pulp stem cells from supernumerary teeth. At passage 3, the cell population showed positive expression of CD 73, CD90 and STRO-1, lacked expression of CD 34 and CD 45. At passage 10, CD 73, CD 90 and STRO-1 showed positive expression while CD 34 and CD 45 showed negative expression. This study indicated that dental pulp stem cells of supernumerary teeth had the properties of mesenchymal stem cells at both early and late passage. Impacted supernumerary teeth could be considered as a noble source of stem cells because of rapid growth and maintaining characteristics of stem cells until late passage.

이 연구의 목적은 발치한 매복 과잉치에서 얻은 치수 유래 세포의 줄기세포 특성을 유세포 분석을 통해 알아보는 것이다. 15명의 환자로부터 채취한 정중 과잉치의 치수 세포를 계대 배양하였고, 3계대와 10계대 세포들을 유세포 분석을 이용하여 분석하였다. 간엽성 줄기세포 표지자 관찰에 사용된 항체 표지자(CD)는 양성 표지자로는 CD 73, CD 90, 그리고STRO-1 와 음성 표지자로는 CD 34, CD 45 이었다. 3계대에서 CD 73, CD 90은 각각 94.82%, 98.86%의 양성반응을, CD 34, CD 45는 각각 2.25%, 2.52%로 음성 반응을 보였으며, STRO-1은 20.93%를 나타냈다. 10계대에서는 CD 73, CD 90은 각각 96.62%, 98.61%의 발현을 보였지만, CD 34, CD 45는 각각 3.86%, 4.14%를 나타냈다. STRO-1은 35.62%로 발현되었다. 이상의 결과에서 과잉치 치수 유래 세포는 간엽성 줄기세포의 특성을 가지며, 3계대와 10계대 모두에서 간엽성 줄기세포의 특성을 유지하고 있다고 사료된다. 이에 빠른 성장 속도와 늦은 계대까지 유지되는 줄기세포능을 고려할 때, 치아 유래 줄기세포의 공여부로서 매복 과잉치의 충분한 활용 가능성을 확인하였다.

Keywords

References

  1. Kim J : Characterization of differentiation of the supernumerary dental pulp stem cells toward the odontoblast by application period of additives. J Korean Acad Pediatr Dent, 42:312-318, 2015.
  2. Evans MJ, Kaufman MH : Establishment in culture of pluripotential cells from mouse embryos. Nature, 292:154-156, 1981. https://doi.org/10.1038/292154a0
  3. Gronthos S, Mankani M, Shi S, et al. : Postnatal human dental pulp stem cells (DPSCs) in vitro and in vivo. Proc Natl Acad Sci USA, 97:13625-13630, 2000. https://doi.org/10.1073/pnas.240309797
  4. Miura M, Gronthos S, Shi S, et al. : SHED: stem cells from human exfoliated deciduous teeth. Proc Natl Acad Sci USA, 100:5807-5812, 2003. https://doi.org/10.1073/pnas.0937635100
  5. Seo BM, Miura M, Shi S, et al. : Investigation of multipotent postnatal stem cells from human periodontal ligament. Lancet, 364:149-155, 2004. https://doi.org/10.1016/S0140-6736(04)16627-0
  6. Morsczeck C, Gotz W, Hoffmann KH, et al. : Isolation of precursor cells (PCs) from human dental follicle of wisdom teeth. Matrix Biol, 24:155-165, 2005. https://doi.org/10.1016/j.matbio.2004.12.004
  7. Sonoyama W, Liu Y, Shi S, et al. : Mesenchymal stem cellmediated functional tooth regeneration in swine. PLoS One, 1:e79, 2006. https://doi.org/10.1371/journal.pone.0000079
  8. Kim J : Managing Complications Related to Multiple Supernumerary Teeth. J Korean Acad Pediatr Dent, 41:180-186, 2014.
  9. Dominici M, Le Blanc K, Horwitz E, et al. : Minimal criteria for defining multipotent mesenchymal stromal cells. The International Society for Cellular Therapy position statement. Cytotherapy, 8:315-317, 2006. https://doi.org/10.1080/14653240600855905
  10. Song JS, Kim SH, Jung HS, et al. : Characterization of stem cells obtained from the dental pulp and periodontal ligament of deciduous teeth. Tissue Eng Regen Med, 7:575-582, 2010.
  11. Park JW, Song JS, Son HK, et al. : Effect of Storage Media and Duration on Pulpal Cell Viability in Exfoliated Deciduous Teeth. J Korean Acad Pediatr Dent, 41:1-7, 2014.
  12. Kim D, Kim J, Roh S, et al. : A nanoscale ridge/groove pattern arrayed surface enhances adipogenic differentiation of human supernumerary tooth-derived dental pulp stem cells in vitro. Arch Oral Biol, 59:765-774, 2014. https://doi.org/10.1016/j.archoralbio.2014.04.014
  13. Almeida-Porada G, Porada C, Zanjani ED : Adult stem cell plasticity and methods of detection. Rev Clin Exp Hematol, 5:26-41, 2001. https://doi.org/10.1046/j.1468-0734.2001.00027.x
  14. Temple S : Stem cell plasticity-building the brain of our dreams. Nat Rev Neurosci, 2:513-520, 2001. https://doi.org/10.1038/35081577
  15. Pittenger MF, Mackay AM, Marshak DR, et al. : Multilineage potential of adult human mesenchymal stem cells. Science, 284:143-147, 1999. https://doi.org/10.1126/science.284.5411.143
  16. Wagers AJ, Weissman IL : Plasticity of adult stem cells. Cell, 116:639-648, 2004. https://doi.org/10.1016/S0092-8674(04)00208-9
  17. Chen SC, Marino V, Gronthos S, Bartold PM : Location of putative stem cells in human periodontal ligament. J Periodontal Res, 41:547-553, 2006. https://doi.org/10.1111/j.1600-0765.2006.00904.x
  18. Hung SC, Chen NJ, Lo WH, et al. : Isolation and characterization of size-sieved stem cells from human bone marrow. Stem Cells, 20:249-258, 2002. https://doi.org/10.1634/stemcells.20-3-249
  19. Kolf CM, Cho E, Tuan RS : Mesenchymal stromall cells. Biology of adult mesenchymal stem cells: regulation of niche, self-renewal and differentiation. Arthritis Res Ther, 9:204, 2007. https://doi.org/10.1186/ar2116
  20. Gay IC, Chen S, MacDougall M : Isolation and characterization of multipotent human periodontal ligament stem cells. Orthod Craniofac Res, 10:149-160, 2007. https://doi.org/10.1111/j.1601-6343.2007.00399.x
  21. Park JH, Song JS, Lee JH, et al. : Characteristics of stem cells derived from the periodontal ligament of supernumerary teeth. Tissue Eng Regen Med, 8:123-131, 2011.