Enhancing Proliferation of Stem Cells from Human Exfoliated Deciduous Teeth (SHED) through hTERT Expression while Preserving Stemness and Multipotency
- PMID: 38878252
- DOI: 10.1007/s12015-024-10746-y
Enhancing Proliferation of Stem Cells from Human Exfoliated Deciduous Teeth (SHED) through hTERT Expression while Preserving Stemness and Multipotency
Abstract
Background: Stem cells from human exfoliated deciduous teeth (SHED) hold promise in regenerative medicine owing to their multipotent capabilities resembling mesenchymal stem cells (MSCs). Despite their potential, SHED have not been extensively investigated because their limited lifespan and unavailability of cell-lines pose challenges for therapeutic applications. This study investigated the effect of ectopic human telomerase reverse transcriptase (hTERT) expression on SHEDs' proliferation while preserving stemness and genomic integrity.
Methods: Deciduous teeth were collected from children aged 6-10 years. After isolation and characterization, the SHED were transduced with pBabe-puro-hTERT retrovirus to establish SHED cell-line, which was evaluated and compared with pBabe-puro (mock control) for stemness, multipotency and growth attributes through flow cytometry, trilineage differentiation, and growth kinetics. We also estimated hTERT gene expression, genomic integrity, and validated cell-line through STR analysis.
Results: Following hTERT transduction, SHED displayed elevated hTERT gene expression while retaining fibroblast-like morphology and mesenchymal stem cell markers. Moreover, after hTERT transduction cellular shape remained same along with increased replicative lifespan and proliferation potential. SHED-hTERT cells exhibited multi-potency and maintained stemness, as evidenced by surface marker expression and multilineage differentiation. Furthermore, genomic integrity was not affected by hTERT integration, as confirmed by STR analysis and CDKN2A gene assessment.
Conclusion: Ectopic hTERT expression in SHED successfully prolonged their replicative lifespan and improved their ability to proliferate and migrate, while preserving their stemness, multipotency and genomic integrity, suggesting minimal carcinogenic risk. Establishment of SHED cell-line holds potential in regenerative medicine applications, especially in cell-based drugs and tissue engineering experiments.
Keywords: Cell-line; Immortalization; SHED; Stem Cells from Human Exfoliated Deciduous Teeth; hTERT Expression.
© 2024. The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature.
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References
-
- Miura, M., Gronthos, S., Zhao, M., Lu, B., Fisher, L. W., Robey, P. G., & Shi, S. (2003). SHED: Stem cells from human exfoliated deciduous teeth. Proceedings of the National Academy of Sciences of the United States of America, 100(10), 5807–5812. https://doi.org/10.1073/PNAS.0937635100/ASSET/C629822E-B3FE-4064-8133-F9...
-
- Yadav, P., Vats, R., Bano, A., Namdev, R., & Bhardwaj, R. (2023). Ameliorative potential of stem cells from human exfoliated deciduous teeth (SHED) in preclinical studies: A meta-analysis. Regenerative Therapy, 24, 117–134. https://doi.org/10.1016/j.reth.2023.06.004 - DOI - PubMed - PMC
-
- Alge, D. L., Zhou, D., Adams, L. L., Wyss, B. K., Shadday, M. D., Woods, E. J., & Goebel, W. S. (2010). Donor-matched comparison of dental pulp stem cells and bone marrow-derived mesenchymal stem cells in a rat model. Journal of Tissue Engineering and Regenerative Medicine, 4(1), 73–81. https://doi.org/10.1002/term.220 - DOI - PubMed - PMC
-
- Liu, J., Ding, Y., Liu, Z., & Liang, X. (2020). Senescence in mesenchymal stem cells: Functional alterations, Molecular mechanisms, and rejuvenation strategies. Frontiers in Cell and Developmental Biology. https://doi.org/10.3389/FCELL.2020.00258/FULL . 8. - DOI - PubMed - PMC
-
- Zhou, X., Hong, Y., Zhang, H., & Li, X. (2020). Mesenchymal stem cell senescence and rejuvenation: Current Status and challenges. Frontiers in Cell and Developmental Biology. https://doi.org/10.3389/FCELL.2020.00364/FULL . 8. - DOI - PubMed - PMC
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