| Project/Area Number |
16209024
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| Research Category |
Grant-in-Aid for Scientific Research (A)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Gastroenterology
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| Research Institution | KEIO UNIVERSITY |
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Principal Investigator |
HIBI Toshifumi Keio University, School of Medicine, Professor, 医学部, 教授 (50129623)
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| Co-Investigator(Kenkyū-buntansha) |
WATANABE Mamoru Tokyo Medical & Dental University, Dept of Medicine, Professor, 医学部, 教授 (10175127)
岡野 栄之 慶應義塾大学, 医学部, 教授 (60160694)
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| Project Period (FY) |
2004 – 2006
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| Project Status |
Completed (Fiscal Year 2006)
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| Budget Amount *help |
¥50,180,000 (Direct Cost: ¥38,600,000、Indirect Cost: ¥11,580,000)
Fiscal Year 2006: ¥8,840,000 (Direct Cost: ¥6,800,000、Indirect Cost: ¥2,040,000)
Fiscal Year 2005: ¥18,070,000 (Direct Cost: ¥13,900,000、Indirect Cost: ¥4,170,000)
Fiscal Year 2004: ¥23,270,000 (Direct Cost: ¥17,900,000、Indirect Cost: ¥5,370,000)
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| Keywords | intestinal epithelial cell / stem cell / musashi-1 / inflammatory bowel disease / regeneration of mucosa / β-catenin / WNT |
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| Research Abstract |
Epithelial renewal occurs in the crypts through a coordinated series of events involving proliferation, differentiation and migration toward the intestinal lumen. It was believed that pluripotent stem cells were located at the lower third of the crypt and generated differentiated cells, such as intestinal epithelial cells, enteroendocrine cells, paneth cells and goblet cells. However, their manner of proliferation and differentiation remains unknown because they lack specific markers and methods of purification and culture. In this study, we propose a new technique to enrich epithelial stem cells and evaluate their phenotype.
We have separated crypts and villi from adult murine small intestine, and have shown that dephosphorylated β catenin and musashi-1 was enriched in crypts fraction. Crypt epithelial cells contained increased numbers of side population (SP) cells and as compared with vinous epithelial cells. In adult intestinal epithelial cells, SP cells have higher amounts of ABCG-2 as compared with main population (MP) cells. Fetal intestinal epithelial cells contained large numbers of SP cells. Dephosphorylated f3 catenin was enriched in SP cells as compared with MP cells.
Then, we focused on WNT/β catenin signaling pathway, and generated TOPGFP mice in which this signaling pathway could be visualized. In these mice, TOPGFP high cells expressed immature markers such as Musashi-1, c-myc. We sorted GFP+ cells from GFP-cells using flow cytometry and compared the expression of various markers in these populations using microarray. Several genes were found to be expressed differentially in GFP+ cells and GFP-cells. These results suggested that TOPGFP high cells were putative stem cells of intestinal epithelia.
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