Preferential enrichment and characterization of human dental pulp stem cell via a genetic engineering-based approach
| 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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Report
(5 results)
Research Products
(22 results)
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[Journal Article] The piggyBac-Based Gene Delivery System Can Confer Successful Production of Cloned Porcine Blastocysts with Multigene Constructs.2016
Author(s)
Sato M, Maeda K, Koriyama M, Inada E, Saitoh I, Miura H, Ohtsuka M, Nakamura S, Sakurai T, Watanabe S, Miyoshi K.
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Journal Title
Int J Mol Sci.
Volume: 17
Issue: 9
Pages: 1424-1424
DOI
NAID
Related Report
Peer Reviewed / Open Access
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[Journal Article] Choice of feeders is important when first establishing iPSCs derived from primarily cultured human deciduous tooth dental pulp cells.2015
Author(s)
Saitoh I, Inada E, Iwase Y, Noguchi H, Murakami T, Soda M, Kubota N, Hasegawa H, Akasaka E, Matsumoto Y, Oka K, Yamasaki Y, Hayasaki H, Sato M
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Journal Title
Cell Medicine
Volume: 8
Issue: 1-2
Pages: 9-23
DOI
Related Report
Peer Reviewed / Open Access
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[Presentation] piggyBac-transposon-mediated gene-delivery efficiently generates stable transfectants from HDDPCs and HDDPC-derived-iPSCs2015
Author(s)
Soda M, Saitoh I, Inada E, Murakami T, Iwase Y, Kubota N, Sawami T, Matsumoto Y, Yamasaki Y, Hayasaki H, Ohshima H, Sato M
Organizer
IADR
Place of Presentation
Boston (America)
Year and Date
2015-03-11 – 2015-03-14
Related Report
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[Presentation] ヒト乳歯歯髄由来iPS細胞樹立におけるフィーダー細胞選択の重要性2013
Author(s)
村上智哉, 齊藤一誠, 稲田絵美, 岩瀬陽子, 長谷川大子, 窪田直子, 松本祐子, 大島邦子, 岡 暁子, 山崎要一, 早崎治明
Organizer
第５１回日本小児歯科学会
Place of Presentation
岐阜県
Related Report
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