Journal of the korean academy of Pediatric Dentistry (대한소아치과학회지)

Korean Academy of Pediatric Dentistry (대한소아치과학회)
권호 표지
DOI QR코드

DOI QR Code

Effect of Storage Media and Duration on Pulpal Cell Viability in Exfoliated Deciduous Teeth

탈락 유치 내 치수 세포의 보관 용액과 기간에 따른 생존

  • Park, Jiwon (Department of Pediatric Dentistry, College of Dentistry, Yonsei University) ;
  • Song, Jeseon (Department of Pediatric Dentistry, College of Dentistry, Yonsei University) ;
  • Lee, Jaeho (Department of Pediatric Dentistry, College of Dentistry, Yonsei University) ;
  • Kim, Seongoh (Department of Pediatric Dentistry, College of Dentistry, Yonsei University) ;
  • Jeon, Mijeong (Oral Science Research Center, College of Dentistry, Yonsei University) ;
  • Jung, Hansung (Division in Anatomy & Developmental Biology, Department of Oral Biology, College of Dentistry, Yonsei University) ;
  • Son, Heungkyu (Department of Pediatric Dentistry, College of Dentistry, Yonsei University)
  • 박지원 (연세대학교 치과대학 소아치과학교실) ;
  • 송제선 (연세대학교 치과대학 소아치과학교실) ;
  • 이제호 (연세대학교 치과대학 소아치과학교실) ;
  • 김성오 (연세대학교 치과대학 소아치과학교실) ;
  • 전미정 (연세대학교 치과대학 구강과학연구소) ;
  • 정한성 (연세대학교 치과대학 구강생물학교실 내 해부발생학 연구실) ;
  • 손흥규 (연세대학교 치과대학 소아치과학교실)
  • Received : 2013.07.03
  • Accepted : 2013.11.14
  • Published : 2014.02.28
Download PDF
⟨ Previous Next ⟩

Abstract

If it is possible to preserve and culture cells from exfoliated deciduous teeth in a readily available storage medium within each family, more stem cells would be obtained. This research is about the effect of storage media and time on pulpal cell viability of exfoliated deciduous teeth. 330 exfoliated deciduous teeth were randomly divided into 11 groups; fresh group, dry group, groups stored in cell culture medium (2, 4, 7 days each), in milk (2, 4, 7 days each), and in saline (2, 4, 7 days each). Primary culture of pulpal cells was conducted in each group and the success rates were compared by calculating the number of teeth with viable cells. The result of primary culture shows that the success rate decreases as the time of storage gets longer. There was no statistical difference between groups stored in the cell culture medium, milk, and saline for 2 and 4 days. However, the groups stored in milk and saline for 7 days showed dramatic decrease in success rate compared to the group stored in the cell culture medium. In conclusion, exfoliated or extracted deciduous teeth can be used to culture pulpal cells when they are stored in milk and saline for a certain period of time; however obtaining viable pulpal cells becomes harder as the storage time gets longer.

가정에서 유치가 탈락하였을 경우 쉽게 구할 수 있는 저장 용액에 보관한 후, 보관되었던 유치에서 세포를 배양할 수 있다면 지금보다 더 많은 유치로부터 줄기 세포를 추출할 수 있을 것이다. 본 연구는 유치의 치수 세포가 다양한 보관 용액의 종류와 기간에 영향을 받는지 확인해 보고자 하였다. 330개의 탈락 유치를 신선군, 건조군, 세포배양액 2, 4, 7일 보관군, 우유 2, 4, 7일 보관군, 생리식염수 2, 4, 7일 보관군으로 각각 30개씩 무작위로 나누었다. 각 군 간의 유치 치수 조직 세포의 일차 배양을 시행하여 성공한 개수와 실패한 개수를 계산하여 생존 성공률을 비교하였다. 일차 배양 결과 보관 기간이 늘어날수록 일차 배양 시 세포의 생존 성공률이 낮아지는 것으로 나타났다. 2일 보관군과 4일 보관군까지 세포배양액, 우유, 생리식염수 간의 보관 용액에 따른 성공률의 유의할 만한 차이는 없었다. 그러나 7일 보관군에서는 세포배양액에 비해 우유와 생리식염수에 보관한 유치에서 성공률이 유의하게 떨어지는 것으로 나타났다. 유치 발거 후 우유나 생리식염수 등의 보관용액에 일정 기간 보관 후 그 치수 조직을 세포 배양에 사용하는 것이 가능하지만 보관 기간이 길어질수록 세포 획득 가능성이 줄어든다.

Keywords

References

  1. Miura M, Gronthos S, Zhao M, 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
  2. Karaoz E, Dogan BN, Aksoy A, et al.: Isolation and in vitro characterisation of dental pulp stem cells from natal teeth. Histochem Cell Biol, 133:95-112, 2010. https://doi.org/10.1007/s00418-009-0646-5
  3. Song JS, Kim Sh, Kim SO, et al.: Characterization of Stem Cells Obtained from the Dental Pulp and Periodontal Ligament of Deciduous Teeth. Tissue Engineering and Regenerative Medicine, 7, 2010.
  4. Wang X, Sha XJ, Li GH, et al.: Comparative characterization of stem cells from human exfoliated deciduous teeth and dental pulp stem cells. Arch Oral Biol, 57:1231-1240, 2012. https://doi.org/10.1016/j.archoralbio.2012.02.014
  5. Kerkis I, Caplan AI: Stem cells in dental pulp of deciduous teeth. Tissue Eng Part B Rev, 18:129- 138, 2012. https://doi.org/10.1089/ten.teb.2011.0327
  6. Song JS, Kim SO, Kim SH, et al.: In vitro and in vivo characteristics of stem cells derived from the periodontal ligament of human deciduous and permanent teeth. Tissue Eng Part A, 18:2040-2051, 2012. https://doi.org/10.1089/ten.tea.2011.0318
  7. Arora V, Arora P, Munshi AK: Banking stem cells from human exfoliated deciduous teeth (SHED): saving for the future. J Clin Pediatr Dent, 33:289- 294, 2009. https://doi.org/10.17796/jcpd.33.4.y887672r0j703654
  8. Souza BD, Luckemeyer DD, Felippe WT, et al.: Effect of temperature and storage media on human periodontal ligament fibroblast viability. Dent Traumatol, 26:271-275, 2010. https://doi.org/10.1111/j.1600-9657.2010.00886.x
  9. Lin DG, Kenny DJ, Barrett EJ, et al.: Storage conditions of avulsed teeth affect the phenotype of cultured human periodontal ligament cells. J Periodontal Res, 35:42-50, 2000. https://doi.org/10.1034/j.1600-0765.2000.035001042.x
  10. de Souza BD, Luckemeyer DD, Felippe WT, et al.: Effect of milk renewal on human periodontal ligament fibroblast viability in vitro. Dent Traumatol, 28:214-216, 2012. https://doi.org/10.1111/j.1600-9657.2011.01078.x
  11. Huang SC, Remeikis NA, Daniel JC: Effects of longterm exposure of human periodontal ligament cells to milk and other solutions. J Endod, 22:30-33, 1996. https://doi.org/10.1016/S0099-2399(96)80233-0
  12. Alacam T, Gorgul G, Omurlu H, Can M: Lactate dehydrogenase activity in periodontal ligament cells stored in different transport media. Oral Surg Oral Med Oral Pathol Oral Radiol Endod, 82:321-323, 1996. https://doi.org/10.1016/S1079-2104(96)80360-X
  13. Liu ZJ, Zhuge Y, Velazquez OC: Trafficking and differentiation of mesenchymal stem cells. J Cell Biochem, 106:984-991, 2009. https://doi.org/10.1002/jcb.22091
  14. Dominici M, Le Blanc K, Mueller I, 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
  15. Gronthos S, Brahim J, Li W, et al.: Stem cell properties of human dental pulp stem cells. J Dent Res, 81:531-535, 2002. https://doi.org/10.1177/154405910208100806
  16. Shi S, Gronthos S: Perivascular niche of postnatal mesenchymal stem cells in human bone marrow and dental pulp. J Bone Miner Res, 18:696-704, 2003. https://doi.org/10.1359/jbmr.2003.18.4.696
  17. Zannettino AC, Paton S, Arthur A, et al.: Multipotential human adipose-derived stromal stem cells exhibit a perivascular phenotype in vitro and in vivo. J Cell Physiol, 214:413-421, 2008. https://doi.org/10.1002/jcp.21210
  18. Freshney R: Culture of animal cells : a manual of basic technique. 5th ed. edn. 2005.
  19. Huang GT, Sonoyama W, Chen J, Park SH: In vitro characterization of human dental pulp cells: various isolation methods and culturing environments. Cell Tissue Res, 324:225-236, 2006. https://doi.org/10.1007/s00441-005-0117-9
  20. Nakamura S, Yamada Y, Katagiri W, et al.: Stem cell proliferation pathways comparison between human exfoliated deciduous teeth and dental pulp stem cells by gene expression profile from promising dental pulp. J Endod, 35:1536-1542, 2009. https://doi.org/10.1016/j.joen.2009.07.024
  21. Eslaminejad MB, Vahabi S, Shariati M, Nazarian H: In vitro Growth and Characterization of Stem Cells from Human Dental Pulp of Deciduous Versus Permanent Teeth. J Dent (Tehran), 7:185-195, 2010.
  22. Govindasamy V, Abdullah AN, Ronald VS, et al.: Inherent differential propensity of dental pulp stem cells derived from human deciduous and permanentteeth. J Endod, 36:1504-1515, 2010. https://doi.org/10.1016/j.joen.2010.05.006
  23. Spath L, Rotilio V, Alessandrini M, et al.: Explantderived human dental pulp stem cells enhance differentiation and proliferation potentials. J Cell Mol Med, 14:1635-1644, 2010.
  24. Lekic PC, Kenny DJ, Barrett EJ: The influence of storage conditions on the clonogenic capacity of periodontal ligament cells: implications for tooth replantation. Int Endod J, 31:137-140, 1998. https://doi.org/10.1046/j.1365-2591.1998.00138.x
  25. Jo JH, Kim SO, Choi HJ, et al.: A comparative study of preserving ability of human periodontal ligament cells stored in different temperatured storage media. J Korean Acad Pediatr Dent 34:36-42, 2007.
  26. Blomlof L, Otteskog P: Viability of human periodontal ligament cells after storage in milk or saliva. Scand J Dent Res, 88:436-440, 1980.
  27. Lindskog S, Blomlof L, Hammarstrom L: Mitoses and microorganisms in the periodontal membrane after storage in milk or saliva. Scand J Dent Res, 91:465-472, 1983.
  28. Ashkenazi M, Sarnat H, Keila S: In vitro viability, mitogenicity and clonogenic capacity of periodontal ligament cells after storage in six different media. Endod Dent Traumatol, 15:149-156, 1999. https://doi.org/10.1111/j.1600-9657.1999.tb00793.x

Cited by

  1. Characterization of Differentiation of the Supernumerary Dental Pulp Stem Cells toward the Odontoblast by Application Period of Additives vol.42, pp.4, 2015, https://doi.org/10.5933/JKAPD.2015.42.4.312