| Project/Area Number |
18592019
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Morphological basic dentistry
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| Research Institution | Matsumoto Dental University |
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Principal Investigator |
YAMAKI Mariko Matsumoto Dental University, Oral Science, Hard Tissue Research, Assistant Professor (90360221)
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| Co-Investigator(Kenkyū-buntansha) |
OZAWA Hidehiro Graduate School of Oral Science, Hard Tissue Research, Professor (60018413)
NAKAMURA Hiroaki Matsumoto Dental University, Second Department of Anatomy, Professor (50227930)
KOBAYASHI Yasuhiro Graduate School of Oral Science, Hard Tissue Research, Associate Professor (20264252)
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| Project Period (FY) |
2006 – 2007
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| Project Status |
Completed (Fiscal Year 2007)
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| Budget Amount *help |
¥3,670,000 (Direct Cost: ¥3,400,000、Indirect Cost: ¥270,000)
Fiscal Year 2007: ¥1,170,000 (Direct Cost: ¥900,000、Indirect Cost: ¥270,000)
Fiscal Year 2006: ¥2,500,000 (Direct Cost: ¥2,500,000)
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| Keywords | Tooth germ mesenchymal cells / MDU1 / Embryonic stem cells / Induction ability / Epithelium-mesenchymal interactions / Collagen 3D carrier / Teratoma / Feeder cells / キメラ胚様体 |
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| Research Abstract |
We hypothesized that a combined use of ES cells and tooth mesenchymal cells is promising for successful development into tooth germ and/or teeth. Based on this hypothesis, we isolated mesenchymal cells from fetal mice, and established a stable cell line, called MDU1 line. We next in vitro cultured ES cells on MDU1 cells, which were used as feeder cells in the culture. Then we found that the ES cells developed into cells that produced keratin and the MDU1 cells into fibroblast-like cells that produced collagen. We therefore considered the cells thus co-cultured capable of developing into tooth germ and/or teeth. We also found that a mixture of cells co-cultured (ES and MDU1 cells) formed embryoid body (EB)-like spheres, called chimera Ebs. We next transplanted chimera Ebs with 3D scaffold made of type I collagen (called collagen sponge, CS) and transplanted the Ebs into sub-renal capsule in mice. Our close observations with histology and immuno-histochemistry identified extensive calcif
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ication, and significant development of odontoblasts and ameloblasts in the grafts. This identification indicates high potential of these Ebs as a good source for tooth germ and/or teeth, and therefore suggests that the absence of the entire structures is due to a small number of cells transplanted. Based on this suggestion, we plan to transplant a larger number of cells co-cultured to identify whether the cells develop into entire structures of tooth germ and/or teeth.
We have observed during the course of the above experiments that ES cells did not form teratoma when transplanted with CS for 7 weeks, whereas ES cells readily formed teratoma when transplanted alone. This observation is extremely important because teratoma formation is one of the most critical problems in practicing stem cell therapy and no perspective for solving this problem has been reported so far. To obtain a better understanding of this phenomenon, we examined gene expression of ES cells, and identified expression of both undifferentiation genes and a tumor marker gene were significantly suppressed when the ES cells were co-cultured with CS. This suggests that CS can directly suppress teratoma formation, and also implies a good method to suppress teratoma formation from cells that experience specific induction from ES cells. We plan to examine this possibility as well. Less
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