| Project/Area Number |
16390578
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| Research Category |
Grant-in-Aid for Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Surgical dentistry
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| Research Institution | The University of Tokyo |
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Principal Investigator |
HONDA Masaki THE UNIVERSITY OF TOKYO, THE INSTITUTE OF MEDICAL SCIENCE, VISITING RESEARCH ASSOCIATE, 医科学研究所, 寄付研究部門教員 (70361623)
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| Co-Investigator(Kenkyū-buntansha) |
ASAHINA Izumi NAGASAKI UNIVERSTY, GRADUATE SCHOOL OF BIOMEDICAL SCIENCES, PROFESSOR, 大学院・医歯薬学総合研究科, 教授 (30221039)
SAGARA Hiroshi THE UNIVERSITY OF TOKYO, THE INSTITUTE OF MEDICAL SCIENCE, RESEARCH ASSOCIATE, 医科学研究所, 助手 (50145041)
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| Project Period (FY) |
2004 – 2006
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| Project Status |
Completed (Fiscal Year 2006)
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| Budget Amount *help |
¥12,800,000 (Direct Cost: ¥12,800,000)
Fiscal Year 2006: ¥2,300,000 (Direct Cost: ¥2,300,000)
Fiscal Year 2005: ¥4,400,000 (Direct Cost: ¥4,400,000)
Fiscal Year 2004: ¥6,100,000 (Direct Cost: ¥6,100,000)
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| Keywords | AMELOGENIN / DENTAL EPITHELIAL CELLS TISSUE / ENAMEL / ENGINEERING / SCAFFOLD / SUBCULTURE / TOOTH BUD / TOOTH BUD CELLS / 歯乳頭細胞 / 歯髄細胞 / 歯嚢細胞 / 幹細胞 / 細胞培養 / フィーダーレイヤー / 歯胚組織 |
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| Research Abstract |
The growth of cells in vitro can provide useful models for investigating their behaviour and tissue engineering. The establishment of culture technique has taken on a new significance. Here, we have reported two novel things on culture system and tissue-engineered tooth.
Firstly, we examined the culture conditions for dental epithelial cells, dental papilla (pulp cells), and dental follicle cells. Especially, we isolated the dental epithelial cells at early crown formation stage and sub-cultured them in 3T3 feeder layer for up to 20 passages. The long-term culture cells expressed mRNA for amelogenin and ameloblastin, as well as enamelysin (MMP-20), which is tissue-specific gene product unique to ameloblasts. These results show that the system is capable of sustaining the multiplication of dental epithelial cells with the characteristics of ameloblasts. Next, we found the dental pulp stem cells that have a potential for SP cells in dental pulp tissues. Finally, we isolated the clone dental follicle cells by single cell clone technique and expanded them in culture. In addition we examined the condition for dental follicle cell to promote the differentiation into periodontal ligament cell that may have a potential for stem cells.
Second, we have evaluated the potential of subcultured odontogenic epithelial cells to form tooth structures in cell-polymer constructs maintained in vivo. Dental epithelial cells were subcultured on feeder layers of 3T3-J2 cells. The subcultured odontogenic epithelial cells were seeded onto collagen sponge scaffolds in combination with dental papilla cells, and transplanted into athymic rats. After 4 weeks, enamel-dentin-like complex structures were present in the implanted constructs. These results show that our culture system produced differentiating ameloblast-like cells that were able to secrete amelogenin proteins and form enamel-like tissues in vivo.
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