| 研究課題/領域番号 |
21K06400
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| 研究機関 | 沖縄科学技術大学院大学 |
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研究代表者 |
Guillaud Laurent 沖縄科学技術大学院大学, 分子神経科学ユニット, グループリーダー (90596222)
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| 研究期間 (年度) |
2021-04-01 – 2024-03-31
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| キーワード | Aging / ATP / LPS / Neurodegeneration |
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| 研究実績の概要 |
The project is going according to plan without significant problems. The data obtain so far are as followed:
1) From the original 4 human iPSC lines, 3 have been successfully cultured, expanded, and cryopreserved.
2) The cell proliferation rate and viability of iPSC from 20 y/o, 40 y/o and 80 y/o individuals have been analyzed and showed significant reduction in the number of colonies, number of cells and cell viability in older iPSCs compared to younger iPSCs.
3) The intracellular levels of ATP in the various iPSC lines have been measured by in vitro bioluminescence assay and revealed a substantial reduction in ATP levels from 80 y/o iPSCs compared to 20 or 40 y/o iPSCs.
4) Reduction in intracellular levels of ATP has also been confirmed in mouse DRG primary cultures from old animals (52 weeks) compared to younger animals (8 weeks), supporting our observations in human iPSCs.
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| 現在までの達成度 (区分) |
現在までの達成度 (区分)
2: おおむね順調に進展している
理由
The progress of the project is going according to plan and the initial characterization of the effect on aging using iPSCs has been achieve by measuring cell profiliferation, viability and ATP levels.
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| 今後の研究の推進方策 |
I am currently analyzing liquid phase separation by FRAP in both human iPSC lines and mouse DRG neurons, as well as quantifying the number of active mitochondria in old samples versus young samples by confocal imaging using TMRE.
Additionally, I am extracting genomic DNA and mitochondrial DNA from iPSCs and DRGs to analyze their DNA methylation levels and determine a potential correlation between genetic clock, mitochondria activity and ATP production, and protein condensation.
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| 次年度使用額が生じた理由 |
The carry over was done because of delay in obtaining cell lines and some reagents.
The carry over budget will be used to performed DNA methylation assay, ATP measurement and live imaging to complete the remaining experiments. Also to attend meeting for presentation and publication fees.
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