2005 Fiscal Year Final Research Report Summary
Identification and separation of pluripotential tissue stem cells and its application for producing somatic cell nuclear transferred cloned mouse
| 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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| Keywords | amniotic epithelial cells / hepatic function / interferon-? / nuclear transfer / somatic clone animals / embryonic stem cells / RNA interference / knockdown mouse |
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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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Research Products
(44 results)
