| Project/Area Number |
25463192
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Multi-year Fund |
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| Section | 一般 |
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| Research Field |
Orthodontics/Pediatric dentistry
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| Research Institution | Kagoshima University |
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Principal Investigator |
Inada Emi 鹿児島大学, 医歯学域附属病院, 助教 (30448568)
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| Co-Investigator(Kenkyū-buntansha) |
佐藤 正宏 鹿児島大学, 医用ミニブタ・先端医療開発研究センター, 教授 (30287099)
齊藤 一誠 新潟大学, 医歯学系, 准教授 (90404540)
野口 洋文 琉球大学, 医学(系)研究科(研究院), 教授 (50378733)
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| Project Period (FY) |
2013-04-01 – 2017-03-31
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| Project Status |
Completed (Fiscal Year 2016)
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| Budget Amount *help |
¥4,680,000 (Direct Cost: ¥3,600,000、Indirect Cost: ¥1,080,000)
Fiscal Year 2016: ¥650,000 (Direct Cost: ¥500,000、Indirect Cost: ¥150,000)
Fiscal Year 2015: ¥1,170,000 (Direct Cost: ¥900,000、Indirect Cost: ¥270,000)
Fiscal Year 2014: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000)
Fiscal Year 2013: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000)
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| Keywords | 歯髄幹細胞 / PiggyBacシステム / 選択的濃縮 / アルカリフォスファターゼ / アルカリ性フォスファターゼ |
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| Outline of Final Research Achievements |
Alkaline phosphatase (ALP) is a useful marker for stem cells that differentiate into human deciduous tooth dental pulp cells (HDDPCs). In this study, we attempted to enrich for ALP-positive cells from HDDPCs via a genetic engineering-based approach. We employed a PiggyBac (PB)-based gene transfer system, which is known to be effective for the enrichment of transfectants. We found that this system is useful for acquiring transfectants when HDDPCs are transfected with PB vectors carrying cDNA for enhanced green fluorescent protein (EGFP). Subsequently, a PB vector carrying the EGFP cDNA and a puromycin resistance gene under the control of the ALP promoter was introduced into HDDPCs together with a transposase expression vector. Unfortunately, this caused accidental arrest of cell proliferation. To overcome this problem, we successfully created a HDDPC-derived immortalized cell line, which will be used as a source for ALP-positive HDDPCs.
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