2010 Fiscal Year Final Research Report
Demonstration of transdifferentiation of other tissue stem cells into dental pulp stem cells by a pulp biomarker
| Project/Area Number |
20390504
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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 |
Dental engineering/Regenerative dentistry
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| Research Institution | National Institute for Longevity Sciences,NCGG |
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Principal Investigator |
NAKASHIMA Misako National Institute for Longevity Sciences,NCGG, 口腔疾患研究部, 室長 (20207773)
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| Co-Investigator(Kenkyū-buntansha) |
IOHARA Kouichiro 国立長寿医療研究センター, 口腔疾患研究部, 研究員 (60435865)
WATANABE Atsushi 国立長寿医療研究センター, 共同利用推進室, 室長 (90321843)
NAKAMURA Hiroshi 愛知学院大学, 歯学部, 教授 (40064878)
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| Project Period (FY) |
2008 – 2010
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| Keywords | 再生歯学 / 歯髄幹細胞 / プロテオーム / 歯髄再生 |
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| Research Abstract |
The present study aimed at identification of a specific biomarker for dental pulp stem cells by systemic protein analysis by differential proteomic expression profiles of dental pulp stem cells and other tissue stem cells. As a result, a new method should be established to induce the dental pulp stem cells from other stem cells and obtain a new cell source with safety and no invasive method. First of all, dental pulp cells were isolated from the extracted tooth pulp after getting the patient's agreement, and CD105^+ cells were isolated by flow cytometry. Moreover, bone marrow CD105^+ cells and amnion CD105+ cells were isolated from a human bone marrow cells and amnion mesenchymal stem cells. After the protein was extracted from these three types of cells respectively, patterns were compared after silver staining in the two-dimensional electrophoretic gels. Nine protein spots were detected only in pulp CD105^+ cells. The proteins were identified by nano liquid chromatography-mass spectrometry (nano-LC/MS) and database searching. They were excised, and the reduction alkylation and the tryptic digestion were done. FK506 binding protein and unnamed protein product of the intermediate filament, were detected. One of the proteins identified with high confidence in pulp CD105+ cells was vimentin. Expression of vimentin mRNA was highest in pulp tissue compared with other variety of tissues, 18 human main tissues and 2 embryonic tissues. Next, interstitial pulp cells were strongly immunostained and the periodontal ligament cells were weakly immunostained with vimentin. All the regenerated tissues after transplantation of the dental pulp stem cells into root canal were similarly immunostained with vimentin. Thus, vimentin can serve as an useful marker for pulp regeneration.
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