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Increased SOX2 expression in three-dimensional sphere culture of dental pulp stem cells KCI 등재후보
Mesenchymal stem cells in the dental pulp exhibit a tendency for differentiation into various dental lineages and hold great potential as a major conduit for regenerative treatment in dentistry. Although they can be readily isolated from teeth, the exact characteristics of these stem cells have not been fully understood so far. When compared to twodimensional (2D) cultures, three-dimensional (3D) cultures have the advantage of enriching the stem cell population. Hence, 3D-organoid culture and 3D-sphere culture were applied to dental pulp cells in the current study. Although the establishment of the organoid culture proved unsuccessful, the 3D-sphere culture readily initiated the stable generation of cell aggregates, which continued to grow and could be passaged to the second round. Interestingly, a significant increase in SOX2 expression was detected in the 3D-spheroid culture compared to the 2D culture. These results indicate the enrichment of the stemness-high population in the 3D-sphere culture. Thus, 3D-sphere culture may act as a link between the conventional and 3D-organoid cultures and aid in understanding the characteristics of dental pulp stem cells.
Materials and Methods
1. Cell culture
2. Isolation and culture of primary dental pulp stemcells
3. Organoid culture of primary dental pulp cells
4. Sphere culture of DPSCs and pDPSCs
5. Immunofluorescence staining
6. RNA isolation and quantitative reverse transcriptionpolymerase chain reaction (RT-PCR)
Results
1. Organoid culture of human DPCs
2. Sphere formation of pDPSCs in serum-freesuspension culture
3. Sphere formation of conventional DPSCs
4. pDPSC Spheres and DPSC spheres express SOX2
Discussion
Acknowledgements
Conflicts of Interest
References
