| Project/Area Number |
15580251
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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 |
Applied animal science
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| Research Institution | Kinki University |
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Principal Investigator |
MITANI Tasuku Kinki University, Institute of Advanced Technology, Associate Professor, 先端技術総合研究所, 助教授 (10322265)
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| Co-Investigator(Kenkyū-buntansha) |
SAEKI Kazuhiro Kinki University, School of Biology-Oriented Science and Technology, Professor, 生物理工学部, 教授 (10298937)
HOSOI Yoshihiko Kinki University, School of Biology-Oriented Science and Technology, Professor, 生物理工学部, 教授 (70192739)
MATSUMOTO Kazuya Kinki University, School of Biology-Oriented Science and Technology, Professor (20298938)
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| Project Period (FY) |
2003 – 2005
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| Project Status |
Completed (Fiscal Year 2005)
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| Budget Amount *help |
¥2,600,000 (Direct Cost: ¥2,600,000)
Fiscal Year 2005: ¥700,000 (Direct Cost: ¥700,000)
Fiscal Year 2004: ¥700,000 (Direct Cost: ¥700,000)
Fiscal Year 2003: ¥1,200,000 (Direct Cost: ¥1,200,000)
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| Keywords | amniotic epithelial cells / hepatic function / interferon-? / nuclear transfer / somatic clone animals / embryonic stem cells / RNA interference / knockdown mouse / 精原細胞 / フローサイトメトリー / 磁気細胞分離システム / α6インテグリン / RNAi / ES細胞 / ALS / 組織幹細胞 / 主要組織適合性抗原 / 細胞移植 / 羊膜細胞 / 単為発生胚 |
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| Research Abstract |
In order to develop a novel method to produce genetically-modified food animals, we aimed to separate somatic stem cells in various tissues applicable for donor cells for producing cloned mice. We also examined an establishment of ES cells from nuclear transferred embryos and their ability to differentiate. Moreover, we examined manipulating an expression of certain target gene.
1.Multipotential amniotic epithelial cells (AECs) were cultured and characterized their properties. AECs showed moderate expression of MHC class I and II and the expression of those molecules by IFN-? was only slightly induced. It was notably that AECs expressed some major hepatic genes. Spermatogonial stem cells (SSC) were enriched from testes using MACS for CD9 or ?6-integrin. SSCs could be cultured in vitro for more than 2 months with expression of CD9, ?6-integrin and Oct4 genes.
2.As aire-deficient mice showed insufficiency of gametogenesis, the expression of aire was examined in gonads and ES cells. AIRE proteins were partially distributed in immature thymus, ovary and ES cells and localized in the nuclei forming nuclear dots.
3.AECs from EGFP-transgenic mice were examined for nuclear transfer. Using the reconstituted embryos which developed to the blastocyst stage, ES cells were established. ES cells were also established from parthenotes and were confirmed their ability to differentiate in vitro and in vivo.
4.Embryonic fibroblasts, cumulus cells, ES cells and AECs were used for producing cloned mouse. Cloned mouse derived from cumulus cells was produced. Factors affecting developmental ability of reconstituted embryos and conditions for improvement of their development were investigated.
5.RNAi-expression vector for SOD1, mutant protein of which induces ALS, was introduced into ES cells and using its stable transfectant, knockdown mice for SOD1 were produced. By crossing this SOD1-siRNA transgenic mouse with human ALS model mouse, siRNA prevented the development ALS.
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