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KAKEN_15K06900seika.pdf
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Title |
Title |
リプログラミング技術を利用した細胞老化のエピゲノム解析
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リプログラミング ギジュツ オ リヨウシタ サイボウ ロウカ ノ エピゲノム カイセキ
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Ripuroguramingu gijutsu o riyōshita saibō rōka no epigenomu kaiseki
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Epigenetic analysis of cellular senescence using the technology for reprogramming cells
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三好, 浩之
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ミヨシ, ヒロユキ
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Miyoshi, Hiroyuki
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慶應義塾大学・医学部 (信濃町) ・特任准教授
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Research team head
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科研費研究者番号 : 70219830
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吉田, 尚美
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ヨシダ, ナオミ
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Yoshida, Naomi
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Collaborator
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楊, 正博
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ヨウ, マサヒロ
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Yō, Masahiro
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Collaborator
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2019
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科学研究費補助金研究成果報告書
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2018
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Abstract |
分裂停止し老化した正常ヒト線維芽細胞からiPS細胞を樹立できるかどうかを検討した。その結果、山中リプログラミング4遺伝子 (Oct3/4, Sox2, Klf4, c-Myc) のみの導入では樹立することはできず、Lin28、SV40LTと2種類のエピジェネティクス制御化合物の追加により、非常に低い頻度ではあるが樹立することができた。老化細胞由来iPS細胞の増殖や形態は、50回以上継代を重ねても、老化前の細胞由来iPS細胞と変わらず、網羅的遺伝子発現、ゲノムワイドのDNAメチル化およびヒストン修飾の比較解析においても差はほとんど見られなかった。
We asked whether iPS cells could be generated from replicatively senescent human fibroblasts. We were not able to generate iPS cells with the four Yamanaka reprogramming factors (Oct3/4, Sox2, Klf4, and c-Myc). However, we succeeded in generation of iPS cells, albeit with extremely low efficiency, using lentiviral vectors expressing the four factors plus Lin28 and non-integrating lentiviral vector for transient expression of SV40LT together with two chemical compounds targeting epigenetic enzymes. There was no difference between iPS cells from senescent cells and control iPS cells for cell growth and morphology up to passage 50 as well as global gene expression profiles, DNA methylation status, and histone modifications. There was also no difference for cellular senescence of fibroblasts differentiated from iPS cells. These results indicate that cellular senescence caused by epigenetic changes can be rejuvenated by reprogramming.
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研究種目 : 基盤研究 (C) (一般)
研究期間 : 2015~2018
課題番号 : 15K06900
研究分野 : 細胞生物学
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