| Project/Area Number |
17390509
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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 |
Conservative dentistry
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| Research Institution | National Institute for Longevity Sciences, National Center for Geriatrics and Gerontology |
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Principal Investigator |
NAKASHIMA Misako National Institute for Longevity Sciences, Lab.of Oral Disease Research, Chief, (研究所)・口腔疾患研究部, 室長 (20207773)
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| Co-Investigator(Kenkyū-buntansha) |
MATSUSHIMA Kenji National Institute for Longevity Sciences, Lab.of Oral Disease Research, Head, (研究所)・口腔疾患研究部, 部長 (90253898)
HASHIMOTO Naohiro National Institute for Longevity Sciences, Dept.of Regenerative Medicine, Head, (研究所)・再生再建医学研究部, 部長 (00208456)
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| Project Period (FY) |
2005 – 2006
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| Project Status |
Completed (Fiscal Year 2006)
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| Budget Amount *help |
¥15,900,000 (Direct Cost: ¥15,900,000)
Fiscal Year 2006: ¥1,400,000 (Direct Cost: ¥1,400,000)
Fiscal Year 2005: ¥14,500,000 (Direct Cost: ¥14,500,000)
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| Keywords | pulp stem cells / amniotic stem cells / dentin regeneration / Side population (SP) cells / flow cytometry / laser microdissection / immunodeficiency / vasculogenesis / angiogenesis / Side population cells |
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| Research Abstract |
The primary amniotic cells from mouse E14 embryo were separated by digestion with dispase and collagenase. CD44^+ ; Sca1^+ cells were further isolated by flow cytometry and expanded on feeders. Those CD44^+; Sca1^+ cells strongly expressed embryonic and adult stem cell markers, POU/Oct4, Sox2, Tert, Rex1, nanog. We had previously isolated side population (SP) cells from porcine dental pulp, which were enriched for stem cell properties. The mRNA expression of the endothelial cell markers, CD31, CD105, Vgfr2 were much stronger in SP cells compared with non-SP cells. To isolate further distinct populations, flow cytometric analyses of the SP cells were performed using antibodies against CD31 and CD146. The CD31^-population was almost CD146^-and represented 50% of total SP cells. The CD31^+ populatio contained both CD146^-and CD146^+ cells, representing 2% and 48% of total SP cells, respectively. CD31^- ; CD146^-SP cells have high vasculogenic potential as well as multi-differentiation potential including odontogenic potential, highly proliferative and migration activities. The subtraction microarray analysis and laser microdissection analysis showed that thyrotropin-releasing hormone degrading ectoenzyme (TRHDE) and low density lipoprotein-related protein 1B (LRP1B) were expressed highly in the pulp stem cells but not in the amniotic stem cells, suggesting possible markers for transdifferentiation of amniotic stem cells. We tried to transdifferentiate the amniotic stem cells into the odontoblastic lineage by some gene electrotransfection, analyzing by expression of TRHDE and differentiation markers for odontoblasts.
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