대한소아치과학회지 (Journal of the korean academy of Pediatric Dentistry)

  • 제45권4호
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  • Pages.492-498
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  • 2018
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  • 1226-8496(pISSN)
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  • 2288-3819(eISSN)
대한소아치과학회 (Korean Academy of Pediatric Dentistry)
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과잉치 치수유래 줄기세포의 장기 배양 특성

Characteristics of Supernumerary Tooth-derived Pulp Cells during Long-term Culture

  • 맹현수 (단국대학교 치과대학 소아치과학교실) ;
  • 김종빈 (단국대학교 치과대학 소아치과학교실) ;
  • 김종수 (단국대학교 치과대학 소아치과학교실)
  • Maeng, Hyunsoo (Department of Pediatric Dentistry, School of Dentistry, Dankook University) ;
  • Kim, Jongbin (Department of Pediatric Dentistry, School of Dentistry, Dankook University) ;
  • Kim, Jongsoo (Department of Pediatric Dentistry, School of Dentistry, Dankook University)
  • 투고 : 2018.06.21
  • 심사 : 2018.07.05
  • 발행 : 2018.11.30
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초록

이 연구는 과잉치 치수로부터 얻은 세포를 더 이상 증식하지 못한다고 판단될 때까지 계대배양을 시행하면서 초기, 중기, 후기 계대의 특성을 비교 분석하였다. 2명의 건강한 6세 남아로부터 3개의 과잉치를 발거하여 치수조직에서 줄기세포를 얻었다. 이를 SNT1(supernumerary tooth 1), SNT2라고 하고, 다른 아이로부터 얻은 과잉치는 SNT3로 명명하였다. SNT1과 SNT2는 똑 같은 시간으로 계대배양 되었고 SNT3은 조금 빠르게 계대배양 되었다. 전체 계대배양의 평균 기간은 $3.6{\pm}1.1$일이었다. 총 $23.3{\pm}0.6$계대까지 배양되었으며, 83일이 소요되었다. 이를 계대를 기준으로 3등분하여 세 군으로 나누었다. I군에서 II군까지 계대배양에 소요된 시간 증가율은 11.90%였으나, II군에서 III군 사이의 증가율은 28.62%로 2.4배가 증가하였다. 22계대까지의 배양기간에 대한 소요 시간을 회귀분석한 결과 y = 0.1169x + 2.25 ($r^2=0.4778$)과 y = 0.1169x + 2.0 ($r^2=0.6444$)으로 추론되었다. 과잉치 치수유래 줄기세포의 정상 배양은 20계대까지 가능할 것으로 판단되었으며, 후기로 넘어가면 계대배양 시간이 2.4배 증가였다. 후기 계대에서는 세포내 부유물이 증가하였고, 형태도 선형으로 변형되는 양상이 관찰되었다.

This study was conducted to investigate the characteristics of subculture times in the early, middle, and late passages by measuring the time under subculture until it was judged that the supernumerary tooth-derived pulp stem cells (sDPSCS) were no longer proliferating. Three supernumerary teeth from two healthy six-years old boys were extracted and stem cells were obtained from the pulp tissue. This was called SNT1 (supernumerary tooth 1), SNT2, and the supernumerary tooth from another child was named SNT3. SNT1 and 2 were subcultured at the same time and SNT3 was subcultured a little faster. The mean time of complete subculture was $3.6{\pm}1.1$ days. Total passages were cultured up to $23.3{\pm}0.6$ and took 83 days. These were divided into three groups based on the passage. The increase rate of time taken in subculture between group I and group II was 11.9%, but the rate between group II and group III was 28.6%, which was 2.4 times increased. The time taken between passages during long-term subculture up to 22 passages shows a regressive pattern y = 0.1169x + 2.25 and y = 0.1169x + 2.0. In conclusion, the passage time of SPSCs increased in late passages, and it shows a similar pattern.

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참고문헌

  1. Evans MJ, Kaufman MH : Establishment in culture of pluripotential cells from mouse embryos. Nature, 292:154-156, 1981. https://doi.org/10.1038/292154a0
  2. Hall PA, Watt FM : Stem cells: the generation and maintenance of cellular diversity. Development, 106:619-633, 1989.
  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 U S A, 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 U S A , 100:5807-5812, 2003. https://doi.org/10.1073/pnas.0937635100
  5. Huang AH, Chen YK, Chan AW, et al. : Isolation and characterization of dental pulp stem cells from a supernumerary tooth. J Oral Pathol Med, 37:571-574, 2008. https://doi.org/10.1111/j.1600-0714.2008.00654.x
  6. Morsczeck C, Moehl C, Hoffmann KH, et al. : In vitro differentiation of human dental follicle cells with dexamethasone and insulin. Cell Biol Int, 29:567-575, 2005. https://doi.org/10.1016/j.cellbi.2005.03.020
  7. Seo BM, Miura M, Shi S, et al. : Recovery of stem cells from cryopreserved periodontal ligament. J Dent Res, 84:907-912, 2005. https://doi.org/10.1177/154405910508401007
  8. 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
  9. Sonoyama W, Liu Y, Huang GT, et al. : Characterization of the apical papilla and its residing stem cells from human immature permanent teeth: a pilot study. J Endod, 34:166-171, 2008. https://doi.org/10.1016/j.joen.2007.11.021
  10. Baek JA, Kim HS, Yoon B, et al. : Efficient culture method for early passage hESCs after Thawing. Korean J Reprod Med, 36:311-319, 2009.
  11. Hayflick L : The limited in vitro lifetime of human diploid cell strains. Exp Cell Res, 37:614-636, 1965. https://doi.org/10.1016/0014-4827(65)90211-9
  12. Park YB, Kim YY, Moon SY, et al. : Alterations of proliferative and differentiation potentials of human embryonic stem cells during long-term culture. Exp Mol Med, 40:98-108, 2008. https://doi.org/10.3858/emm.2008.40.1.98
  13. Bruder SP, Jaiswal N, Haynesworth SE : Growth kinetics, selfrenewal, and the osteogenic potential of purified human mesenchymal stem cells during extensive subcultivation and following cryopreservation. J Cell Biochem, 64:278-294, 1997. https://doi.org/10.1002/(SICI)1097-4644(199702)64:2<278::AID-JCB11>3.0.CO;2-F
  14. Shin Y, Kim J, Kim J : Relationship with Passage Time of Human Dental Pulp Stem Cells from Supernumerary Tooth by Classification. J Korean Acad Pediatr Dent, 43:419-426, 2016.
  15. Shin J, Kim J : Comparison of Mineralization in Each Passage of Dental Pulp Stem Cells from Supernumerary Tooth. J Korean Acad Pediat Dent, 44:350-357, 2017.
  16. Abo-Aziza FAM, A A Z : The Impact of Confluence on Bone Marrow Mesenchymal Stem (BMMSC) Proliferation and Osteogenic Differentiation. Int J Hematol Oncol Stem Cell Res, 11:121-132, 2017.
  17. Stenderup K, Justesen J, Clausen C, Kassem M : Aging is associated with decreased maximal life span and accelerated senescence of bone marrow stromal cells. Bone, 33:919-926, 2003. https://doi.org/10.1016/j.bone.2003.07.005
  18. Min JH, Ko SY, Jang YJ, et al. : Dentinogenic potential of human adult dental pulp cells during the extended primary culture. Hum Cell, 24:43-50, 2011. https://doi.org/10.1007/s13577-011-0010-7
  19. Park S, Kang HM, Kim H, et al. : Characterization of umbilical cord-derived stem cells during expansion in vitro. Korean J Reprod Med, 36:23-34, 2009.
  20. Ha SY, Jee BC, Moon SY, et al. : Cryopreservation of human embryonic stem cells without the use of a programmable freezer. Hum Reprod, 20:1779-1785, 2005. https://doi.org/10.1093/humrep/deh854
  21. Rosler ES, Fisk GJ, Carpenter MK, et al. : Long-term culture of human embryonic stem cells in feeder-free conditions. Dev Dyn, 229:259-274, 2004. https://doi.org/10.1002/dvdy.10430
  22. Draper JS, Smith K, Andrews PW, et al. : Recurrent gain of chromosomes 17q and 12 in cultured human embryonic stem cells. Nat Biotechnol, 22:53-54, 2004. https://doi.org/10.1038/nbt922
  23. Bonab MM, Alimoghaddam K, Nikbin B, et al. : Aging of mesenchymal stem cell in vitro. BMC Cell Biol, 7:14, 2006. https://doi.org/10.1186/1471-2121-7-14