Análisis del efecto del suplemento FREEADD en la morfología y diferenciación osteogénica de células troncales de pulpa dental humana
Palabras clave:
celulas dentales, celulas troncales de la pulpa dental, suero fetal bovino, FreeAdd, osteogenesis
Resumen
El cultivo celular es una técnica esencial en la investigación biológica, que facilita estudios in vitro sobre diversas enfermedades. En este contexto, el suero fetal bovino (SFB) es un suplemento clave, ya que proporciona los nutrientes necesarios para el crecimiento celular. Sin embargo, su producción plantea importantes preocupaciones éticas y la variabilidad entre lotes que puede comprometer la reproducibilidad de los resultados experimentales. En respuesta a estas limitaciones, varias compañías han desarrollado medios definidos químicamente y libres de suero, como FreeAdd. Debido a ello, en este estudio se evaluó el efecto del FreeAdd en la morfología, cambios en la expresión de los genes de pluripotencia y capacidad de diferenciación osteogénica de las células troncales de pulpa dental humana (CTPDh). Utilizando el medio de cultivo convencional (Medio Eagle’s alfa modificado) y un medio comercial para el uso del reemplazo de suero (F10 Ham's) suplementados con el SFB (control) o con el FreeAdd. Los resultados obtenidos mostraron una morfología celular más corta cuando se encontraban en presencia de FreeAdd a comparación de las que se encontraban en SFB que era fusiforme. A pesar de ello, no hubo cambios en los niveles de transcritos de genes de pluripotencia, sin embargo, bajo las condiciones de suplementación empleadas en este trabajo, la diferenciación osteogénica fue menor cuando se encontraban en presencia de FreeAdd.
Citas
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Yuan, J., Liu, X., Chen, Y., Zhao, Y., Liu, P., Zhao, L., & Han, W. (2017). Effect of SOX2 on osteogenic differentiation of dental pulp stem cells. Cellular and Molecular Biology (Noisy-Le-Grand, France), 63(1), 41–44. https://doi.org/10.14715/CMB/2017.63.1.8
Chelladurai, K. S., Durairaj, J., Christyraj, S., Rajagopalan, K., Yesudhason, B. V., Venkatachalam, S., Mohan, M., Chellathurai Vasantha, N., Retnaraj, J., & Selvan Christyraj, S. (2017). Alternative to FBS in animal cell culture - An overview and future perspective. Heliyon, e07686. https://doi.org/10.1016/j.heliyon.2021.e07686
Cowper, M., Frazier, T., Wu, X., Curley, J. L., Ma, M. H., Mohiuddin, O. A., Dietrich, M., McCarthy, M., Bukowska, J., & Gimble, J. M. (2019). Human Platelet Lysate as a Functional Substitute for Fetal Bovine Serum in the Culture of Human Adipose Derived Stromal/Stem Cells. Cells, 8(7). https://doi.org/10.3390/CELLS8070724
de Girolamo, L., Sartori, M. F., Albisetti, W., & Brini, A. T. (2007). Osteogenic differentiation of human adipose-derived stem cells: comparison of two different inductive media. Journal of Tissue Engineering and Regenerative Medicine, 1(2), 154–157. https://doi.org/10.1002/TERM.12
Eydt, C., Geburek, F., Schröck, C., Hambruch, N., Rohn, K., Pfarrer, C., & Staszyk, C. (2016). Sternal bone marrow derived equine multipotent mesenchymal stromal cells (MSCs): investigations considering the sampling site and the use of different culture media. Veterinary Medicine and Science, 2(3), 200–210. https://doi.org/10.1002/VMS3.36
Foroutan, T., & Foroutan, T. (2016). International Journal of Organ Transplantation Medicine Comparison of Differentiation of Induced Pluripotent Stem Cells and Bone-Marrow Mesenchymal Stem Cells to Osteoblast: Osteogenesis versus Pluripotency. Int J Organ Transplant Med., 7, 91–96. www.ijotm.com
Gstraunthaler, G. (2003). Alternatives to the use of fetal bovine serum: Serum-free cell culture. ALTEX - Alternatives to Animal Experimentation, 20(4), 275–281. https://doi.org/10.14573/ALTEX.2003.4.257
Hawkes, P. W. (2015). Fetal bovine serum: Geographic origin and regulatory relevance of viral contamination. Bioresources and Bioprocessing, 2(1), 1–5. https://doi.org/10.1186/S40643-015-0063-7/TABLES/1
Kyllönen, L., Haimi, S., Mannerström, B., Huhtala, H., Rajala, K. M., Skottman, H., Sándor, G. K., & Miettinen, S. (2013). Effects of different serum conditions on osteogenic differentiation of human adipose stem cells in vitro. Stem Cell Research and Therapy, 4(1), 1–15. https://doi.org/10.1186/SCRT165/FIGURES/7
Lee, M. S., Youn, C., Kim, J. H., Park, B. J., Ahn, J., Hong, S., Kim, Y. D., Shin, Y. K., & Park, S. G. (2017). Enhanced Cell Growth of Adipocyte-Derived Mesenchymal Stem Cells Using Chemically-Defined Serum-Free Media. International Journal of Molecular Sciences 2017, Vol. 18, Page 1779, 18(8), 1779. https://doi.org/10.3390/IJMS18081779
Liu, L., Ling, J., Wei, X., Wu, L., & Xiao, Y. (2009). Stem Cell Regulatory Gene Expression in Human Adult Dental Pulp and Periodontal Ligament Cells Undergoing Odontogenic/Osteogenic Differentiation. Journal of Endodontics, 35(10), 1368–1376. https://doi.org/10.1016/J.JOEN.2009.07.005
Matsuoka, F., Takeuchi, I., Agata, H., Kagami, H., & Shiono, H. (2013). Morphology-Based Prediction of Osteogenic Differentiation Potential of Human Mesenchymal Stem Cells. PLoS ONE, 8(2), 55082. https://doi.org/10.1371/journal.pone.0055082
Mercado-Rubio, M. D., Pérez-Argueta, E., Zepeda-Pedreguera, A., Aguilar-Ayala, F. J., Peñaloza-Cuevas, R., Kú-González, A., Rojas-Herrera, R. A., Rodas-Junco, B. A., & Nic-Can, G. I. (2021). Similar features, different behaviors: A comparative in vitro study of the adipogenic potential of stem cells from human follicle, dental pulp, and periodontal ligament. Journal of Personalized Medicine, 11(8), 738. https://doi.org/10.3390/JPM11080738/S1
Panella, S., Muoio, F., Jossen, V., Harder, Y., Eibl-Schindler, R., & Tallone, T. (2021). Chemically Defined Xeno- and Serum-Free Cell Culture Medium to Grow Human Adipose Stem Cells. Cells 2021, Vol. 10, Page 466, 10(2), 466. https://doi.org/10.3390/CELLS10020466
Paradkar, P. H., Db Vaidya, A., Talwalkar, S. C., Mishra, L. S., Agashe, S. V, & Vaidya, R. A. (2019). Bovine whey protein and other biological fluids as alternative to fetal bovine serum in supplementing cell culture media. Indian Journal of Experimental Biology, 57, 123–130.
Piletz, J. E., Drivon, J., Eisenga, J., Buck, W., Yen, S., McLin, M., Meruvia, W., Amaral, C., & Brue, K. (2018). Human Cells Grown With or Without Substitutes for Fetal Bovine Serum. Https://Doi.Org/10.1177/2155179018755140, 10, 215517901875514. https://doi.org/10.1177/2155179018755140
Pilgrim, C. R., McCahill, K. A., Rops, J. G., Dufour, J. M., Russell, K. A., & Koch, T. G. (2022). A Review of Fetal Bovine Serum in the Culture of Mesenchymal Stromal Cells and Potential Alternatives for Veterinary Medicine. Frontiers in Veterinary Science, 9, 859025. https://doi.org/10.3389/FVETS.2022.859025
Popov, A., Scotchford, C., Grant, D., & Sottile, V. (2019). Impact of Serum Source on Human Mesenchymal Stem Cell Osteogenic Differentiation in Culture. International Journal of Molecular Sciences, 20(20). https://doi.org/10.3390/IJMS20205051
Thamm, K., Möbus, K., Towers, R., Baertschi, S., Wetzel, R., Wobus, M., & Segeletz, S. (2022). A chemically defined biomimetic surface for enhanced isolation efficiency of high-quality human mesenchymal stromal cells under xenogeneic/serum-free conditions. Cytotherapy, 24(10), 1049–1059. https://doi.org/10.1016/J.JCYT.2022.06.003
van der Valk, J., Brunner, D., De Smet, K., Fex Svenningsen, Å., Honegger, P., Knudsen, L. E., Lindl, T., Noraberg, J., Price, A., Scarino, M. L., & Gstraunthaler, G. (2010). Optimization of chemically defined cell culture media – Replacing fetal bovine serum in mammalian in vitro methods. Toxicology in Vitro, 24(4), 1053–1063. https://doi.org/10.1016/J.TIV.2010.03.016
Verma, A., Verma, M., & Singh, A. (2020). Animal tissue culture principles and applications. Animal Biotechnology: Models in Discovery and Translation, 269–293. https://doi.org/10.1016/B978-0-12-811710-1.00012-4
Witzeneder, K., Lindenmair, A., Gabriel, C., Höller, K., Theiß, D., Redl, H., & Hennerbichler, S. (2013). Human-Derived Alternatives to Fetal Bovine Serum in Cell Culture. Transfus Med Hemother, 40, 417–423. https://doi.org/10.1159/000356236
Yuan, J., Liu, X., Chen, Y., Zhao, Y., Liu, P., Zhao, L., & Han, W. (2017). Effect of SOX2 on osteogenic differentiation of dental pulp stem cells. Cellular and Molecular Biology (Noisy-Le-Grand, France), 63(1), 41–44. https://doi.org/10.14715/CMB/2017.63.1.8
Publicado
2026-04-30
Sección
Artículos de Investigación
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