In Vitro Cultivation of Dental Pulp Stem Cells from Human Exfoliated Deciduous Teeth in Low-Xenogeneic-Serum Containing Media
(Original Article – Experimental Study)
Authors:
T. Suchánková Kleplová 1,2; K. Z. Browne 2; T. Soukup 2; J. Suchánek 1,2
Authors‘ workplace:
Ústav histologie a embryologie LF UK, Hradec Králové
1; Stomatologická klinika LF UK a FN, Hradec Králové
2
Published in:
Česká stomatologie / Praktické zubní lékařství, ročník 116, 2016, 1, s. 3-11
Category:
Original articles
Overview
Introduction and aims:
Recently, the human regenerative medicine has been departing from the use of xenogeneic materials for the possibility of zoonosis transfer and development of prion infection. We have focused our research on cultivation of Dental Pulp Stem Cells from Human Exfoliated Deciduous Teeth (SHED) in a medium with lowered concentration of Fetal Calf Serum (FCS), which decreases the chances of the above mentioned negative impacts on the cultivated cell population. In 2011, Karbanova et al. have shown that the Dental Pulp Stem Cells (DPSCs) from Impacted Third Molars can be viably cultured in a xenogeneic medium containing as little as 2% FCS. Our hypothesis, that SHED should react similarly to isolation and cultivation methods as DPSCs, follows from their similarities in the place of origin and their niches.
Method:
We have tested this hypothesis on three lineages of SHED from varying donours. The SHED were isolated from the exfoliated deciduous teeth dental pulp using enzymatic dissociation and seeded onto cultivation dishes containing the test medium of 2% of FCS. The cells were then continuously expanded over the Hayflick Limit, the proof of “stemness”. Following the proliferation potential test, we have examined the phenotype of the cultured undifferentiated cells which expressed high positivity of the Mesenchymal Stem Cell (MSC) surface markers, which agrees with the characteristic SHED phenotype. After we have audited the impact of the isolation and cultivation methods, we have proceeded to test their differentiation potential by seeding the cultured SHED into various differentiation media. The SHED have successfully differentiated into all the targeted unipotent tissue and cell types, namely: osteoblasts, chondrocytes, endothelocytes, myofibroblasts and neural cells.
Results:
Overall, our data shows that the SHED cultured in low-xenogeneic-serum medium, unlike the SHED cultured in high-xenogeneic-serum media, express lowered positivity of the same surface markers.
Conclusion:
Our results suggest that SHED are exceptionally promising and available mesenchymal stem cell population, especially applicable for neuroregenerative medicine.
Keywords:
stem cells from human exfoliated deciduous teeth – SHED – xenogeneic serum – fetal calf serum, 2% FCS – enzymatic digestion – differentiation potential
Sources
1. Akpinar, G., Kasap, M., Aksoy, A., et al.: Phenotypic and proteo-mic characteristics of human dental pulp derived mesenchymal stem cells from a natal, an exfoliated deciduous, and an impacted third molar tooth. Stem Cells Int., 2014. 2014:457059. doi: 10.1155/2014/457059, 2014.
2. Arora, V., Arora, P.: Pediatric stem cells – the future ahead. Int. J. Biomed. Res., roč. 3, 2012, č. 11, s. 414–421.
3. Bakopoulou, A., Leyhausen, G., Volk, J., et al.: Assessment of the impact of two different isolation methods on the osteo/odontogenic differentiation potential of human dental stem cells derived from deciduous teeth. Calcif. Tissue Int., roč. 88, 2011, č. 2, s. 130–141.
4. Cordeiro, M. M., Dong, Z., Kaneko, T., et al.: Dental pulp tissue engineering with stem cells from exfoliated deciduous teeth. J. Endod., roč. 34, 2008, č. 8, s. 962–969.
5. 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, roč. 8, 2006, č. 4, s. 315–317.
6. Govindasamy, V., et al.: Inherent differential propensity of dental pulp stem cells derived from human deciduous and permanent teeth. J. Endod., roč. 36, 2010, č. 9, s. 1504–1515.
7. Chen Xu, N. K., Shen, Y. Y.: Isolation and identification of stem cells derived from human exfoliated deciduous teeth. Nan Fang Yi Ke Da Xue Xue Bao J. Southern Medical University, roč. 29, 2009, č. 3, s. 479–482.
8. Ishiy, F. A. A., Fanganiello, R. D., Griesi-Oliveira, K., et al.: Improvement of in vitro osteogenic potential through differentiation of induced pluripotent stem cells from human exfoliated dental tissue towards mesenchymal-like stem cells. Stem Cells International, 2015.
9. Karbanová, J., Soukup, T., Suchánek, J., et al.: Characterization of dental pulp stem cells from impacted third molars cultured in low serum-containing medium. Cells Tissues Organ, roč. 193, 2011, č. 6, s. 344–365.
10. Kerkis, I., Kerkis, A., Dozortsev, D., et al.: Isolation and characterization of a population of immature dental pulp stem cells expressing OCT-4 and other embryonic stem cell markers. Cells Tissues Organs, roč. 184, 2006, č. 3–4, s. 105–116.
11. Kerkis, I., Caplan, A. I.: Stem cells in dental pulp of deciduous teeth. Tissue Engineering Part B: Reviews, roč. 18, 2011, č. 2, s. 129–138.
12. Kiyani, M. N., Rabiei, F., Mardani, M., et al.: Induced in vitro differentiation of neural-like cells from human exfoliated deciduous teeth-derived stem cells. Int. J. Dev. Biol., roč. 55, 2011, s. 189–195.
13. Law, C. S.: Management of premature primary tooth loss in the child patient. J. Calif. Dent. Assoc., roč. 48, 2013, s. 612–618.
14. Lindroos, B., Mäenpää, K., Ylikomi, T., et al.: Characterisation of human dental stem cells and buccal mucosa fibroblasts. Biochem. Biophys. Res. Commun., roč. 368, 2008, č. 2, s. 329–335.
15. Macena, M. C. B., Katz, C. R. T., Heimer, M. V., et al.: Space changes after premature loss of deciduous molars among Brazilian children. Am. J. Orthod. Dentofacial Orthop., roč. 140, 2011, č. 6, s. 771–778.
16. Mackay, A. M., Beck, S. C., Murphy, J. M., et al.: Chondrogenic differentiation of cultured human mesenchymal stem cells from marrow. Tissue Eng, roč. 4, 1998, č. 4, s. 415–428.
17. Miura, M., Gronthos, S., Zhao, M., Lu, B., et al.: SHED: stem cells from human exfoliated deciduous teeth. Proc. Natl. Acad. Sci. USA, roč. 100, 2003, č. 10, s. 5807–5812.
18. 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. Endodont, roč. 35, 2009, č. 11, s. 1536–1542.
19. Pittenger, M. F., Mackay, A. M., Beck, S. C., et al.: Multilineage potential of adult human mesenchymal stem cells. Science, 1999, č. 284, s. 143–147.
20. Shahdadfar, A., Frønsdal, K., Haug, T., et al.: In vitro expansion of human mesenchymal stem cells: choice of serum is a determinant of cell proliferation, differentiation, gene expression, and transcriptome stability. Stem Cells, roč. 23, 2005, č. 9, s. 1357–1366.
21. Suchánek, J., Soukup, T., Ivancaková, R., et al.: Human dental pulp stem cells-isolation and long term cultivation. Acta medica (Hradec Králové)/Universitas Carolina, Facultas Medica Hradec Králové, roč. 50, 2007, s. 195–201.
22. Suchánek, J., Visek, B., Soukup, T., et al.: Stem cells from human exfoliated deciduous teeth-isolation, long term cultivation and phenotypical analysis. Acta medica (Hradec Králové)/Universitas Carolina, Facultas Medica Hradec Králové, roč. 53, 2010, č. 2, s. 93–99.
Labels
Maxillofacial surgery Orthodontics Dental medicineArticle was published in
Czech Dental Journal
2016 Issue 1
Most read in this issue
-
Oral Carriage of Staphylococcus aureus among Dental Patients in Dependence on Conditions in Oral Cavity
(Original Article – Retrospective Study) -
Maturogenesis
Part I. Introduction, Stem Cells, Growth Factors and Matrix
(Review) -
In Vitro Cultivation of Dental Pulp Stem Cells from Human Exfoliated Deciduous Teeth in Low-Xenogeneic-Serum Containing Media
(Original Article – Experimental Study)