| Project/Area Number |
26893074
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| Research Category |
Grant-in-Aid for Research Activity Start-up
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| Allocation Type | Single-year Grants |
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| Research Field |
Dental engineering/Regenerative dentistry
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| Research Institution | Tokyo Medical and Dental University |
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Principal Investigator |
Yamamoto Mioko 東京医科歯科大学, 歯学部附属病院, 医員 (50732749)
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| Project Period (FY) |
2014-08-29 – 2016-03-31
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| Project Status |
Completed (Fiscal Year 2015)
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| Budget Amount *help |
¥2,730,000 (Direct Cost: ¥2,100,000、Indirect Cost: ¥630,000)
Fiscal Year 2015: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000)
Fiscal Year 2014: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000)
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| Keywords | 三次元スフェロイド培養 |
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| Outline of Final Research Achievements |
For 3D spheroid culture, mouse dental papilla cells (MDPs) were cultured for 24 hours in 96-well low-attachment culture plates. Flat-bottomed plates were used for two-dimensional (2D) monolayer culture as a control. Following RNA extraction and complementary DNA synthesis from samples, real-time PCR was performed to evaluate the expression of alkaline phosphatase (Alp) and bone morphogenetic protein2 (BMP2) using specific primers. Up-regulation of Alp and Bmp2 expression was observed in the 3D spheroid-cultured MDPs, however, the expression of these genes was low in the 2D monolayer-cultured MDPs. In order to study the effects of integrin signaling, Focal adhesion kinase and Extracellular Signal-regulated Kinase 1/2 inhibitors were added to the 3D-spheroid cultured MDPs. Then, down-regulation of Alp and Bmp2 was observed in the presence of these inhibitors. In conclusion, 3D-spheroid culture induced the odonto-/osteoblastic differentiation of MDPs probably through integrin signaling.
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