| Project/Area Number |
21590803
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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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 | Tokyo Medical and Dental University |
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Principal Investigator |
NAKAMURA Tetsuya 東京医科歯科大学, 大学院・医歯学総合研究科, 寄附講座教員 (70265809)
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| Co-Investigator(Kenkyū-buntansha) |
WATANABE Mamoru 東京医科歯科大学, 医歯学総合研究科, 教授 (10175127)
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| Project Period (FY) |
2009 – 2011
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| Project Status |
Completed (Fiscal Year 2011)
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| Budget Amount *help |
¥4,680,000 (Direct Cost: ¥3,600,000、Indirect Cost: ¥1,080,000)
Fiscal Year 2011: ¥910,000 (Direct Cost: ¥700,000、Indirect Cost: ¥210,000)
Fiscal Year 2010: ¥910,000 (Direct Cost: ¥700,000、Indirect Cost: ¥210,000)
Fiscal Year 2009: ¥2,860,000 (Direct Cost: ¥2,200,000、Indirect Cost: ¥660,000)
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| Keywords | 消化管上皮幹細胞 / ライブセルイメージング / 初代培養技術 / 細胞移植治療 / 大腸上皮幹細胞 / ライブイメージング / 細胞増殖 / 細胞分化 / 初代培養 / 幹細胞移植 / 再生治療 / 炎症性腸疾患 |
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| Research Abstract |
We have developed a novel culture method for colonic epithelial cells that allows long-term maintenance and efficient expansion of stem cells in vitro. Colonic crypts from normal adult mice were isolated and three-dimensionally cultured in serum-free medium supplemented with a combination of growth factors. The crypt cells formed a round cystic organoid consisting of epithelial monolayer of multi-lineage cells, continuously grew with increasing size of organoid structure, and could be propagated for months without losing their properties. Importantly, Lgr5+colonic stem cells were constantly maintained for a long time period in our culture. In order to test transplantability of these cultured cells, GFP+colonic cells were re-introduced into the lumen of mouse colon superficially damaged by DSS treatment. As observed 4 weeks post-transplantation, the infused organoids readily integrated into the epithelium to form self-renewing GFP+epithelial patches that were histologically normal. Similar results were obtained with colon organoids that were derived from a single Lgr5+colon stem cell after extensive in vitro expansion.
The techniques that we have developed would be of significant help to build a basis to develop a novel colon stem cell therapy based on in vitro expansion of a single adult colonic stem cell.
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