Three-dimensional Sphere-forming Cells Are Unique Multipotent Cell Population in Dental Pulp Cells
| Autor: | Nadia Chugal, Ki-Hyuk Shin, Neha Mohindroo, Sung Hee Lee, Charlotte Ellen Martin, Mo K. Kang, Andrew Inaba, Reuben H. Kim, No-Hee Park, Deepika Ganesh |
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| Rok vydání: | 2016 |
| Předmět: |
0301 basic medicine
Population Blotting Western Mice Nude Flow cytometry 03 medical and health sciences Mice Dental pulp stem cells medicine Animals Humans education General Dentistry Cells Cultured Dental Pulp education.field_of_study medicine.diagnostic_test Chemistry Reverse Transcriptase Polymerase Chain Reaction Multipotent Stem Cells Mesenchymal stem cell Cell Differentiation Cell biology Transplantation 030104 developmental biology Phenotype Immunology Pulp (tooth) Alkaline phosphatase Stem cell Transcription Factors |
| Zdroj: | Journal of endodontics. 43(8) |
| ISSN: | 1878-3554 |
| Popis: | Introduction Mesenchymal stem cells (MSCs) are typically cultured as adherent monolayer using a conventional tissue culture technique. However, this technique incompletely reproduces an in vivo microenvironment of stem cells and results in the loss of stemness properties. Three-dimensional (3D) sphere culture is one of the most widely used 3D culture techniques that have been developed to recapitulate the in vivo microenvironment. However, the stemness and multilineage differentiation capacity of spheres derived from dental pulp stem cells (DPSCs) have not been well investigated. Methods DPSCs were cultured and examined for the sphere-forming ability in serum-free, nonadherent conditions. The expression of pluripotency transcription factors was assayed by reverse transcription quantitative polymerase chain reaction and Western blot analysis. The expression of MSC-associated markers was determined by flow cytometry. Multilineage differentiation capacity was examined by alkaline phosphatase, alizarin red S, and oil red O assays. Subcutaneous transplantation in nude mice was used to examine the in vivo mineralized tissue-forming ability of sphere and adherent monolayer cells derived from DPSCs. Results We showed that DPSCs form spheres. DPSC spheres exhibited a distinct stem cell phenotype characterized by robust expression of pluripotency transcription factors and decreased expression of MSC-associated markers compared with their corresponding adherent monolayer cells. Functionally, DPSC spheres exhibited enhanced in vitro multilineage differentiation capacity. The expression of multilineage differentiation-related genes was also highly increased in DPSC spheres. Furthermore, DPSC sphere cells possessed higher in vivo mineralized tissue-forming ability than adherent monolayer cells. Conclusions Our findings indicate that sphere-forming cells are unique multipotent cell populations in DPSCs. Our study further suggests that DPSC spheres may provide a unique opportunity for pulp tissue regeneration. |
| Databáze: | OpenAIRE |
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