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KAKEN_26670432seika.pdf
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Title |
Title |
転写調節因子を自在誘導できるESバンクを用いた腎尿細管ダイレクトリプログラミング
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Kana |
テンシャ チョウセツ インシ オ ジザイ ユウドウデキル ES バンク オ モチイタ ジン ニョウサイカン ダイレクト リプログラミング
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Tensha chosetsu inshi o jizai yudodekiru ES banku o mochiita jin nyosaikan dairekuto ripuroguramingu
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The direct differentiation method of renal tubular cells by master transcription factors identified from TF-inducible human ES bank
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伊藤, 裕
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Kana |
イトウ, ヒロシ
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Romanization |
Ito, Hiroshi
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慶應義塾大学・医学部(信濃町)・教授
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Research team head
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科研費研究者番号 : 40252457
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洪, 実
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コウ, ミノル
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Ko, Minoru
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慶應義塾大学・医学部(信濃町)・教授
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Research team member
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科研費研究者番号 : 50631199
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門川, 俊明
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モンカワ, トシアキ
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Monkawa, Toshiaki
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慶應義塾大学・医学部(信濃町)・教授
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Research team member
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科研費研究者番号 : 80286484
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平塚, 健
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ヒラツカ, ケン
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Hiratsuka, Ken
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慶應義塾大学・医学部(信濃町)・博士課程大学院生
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Research team member
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2016
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科学研究費補助金研究成果報告書
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2015
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Abstract |
我々は, 腎尿細管への分化誘導に関わる転写因子を明らかにするために転写調節因子を自在に誘導できるヒトESバンクのマイクロアレイデータを用いて網羅的解析を行った。複数個の候補となる転写因子が明らかになり, これらを合成mRNAの形でヒトES細胞に導入することで5日間で中間中胚葉のマーカーであるOSR1を初め近位尿細管のマーカーであるAQP1, MEGALINのmRNAレベルでの上昇, さらに, AQP1, LTLの蛋白レベルでの発現が確認できた。今回, 我々は近位尿細管の分化誘導に必須となる転写因子を同定し, 合成mRNAを用いた全く新しい分化誘導方法を示すことに成功した。
To find transcription factors which promote differentiation towards renal tubular cells, we utilize the human ES lines with doxycycline-controllable transcription factors (TF-inducible hES bank) and performed exhaustive search for DNA microarray data. Some candidate transcription factors were identified by in silico analysis. By using the lipofection method, we transfected the synthetic mRNA of target transcription factors to human ES cells, and cultured them. Five days after the transfection, we were able to observe characteristic morphological changes in the differentiated cells. The mRNA expression of OSR1, AQP1, and MEGALIN were increased. Moreover, the protein expression of AQP1 and LTL were also detected in the differentiated cells. We identified specific transcription factors for differentiation toward proximal tubular cells, and demonstrated that the differentiation of proximal tubular cell phenotype from human ES cells by a novel method using synthetic mRNA.
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研究種目 : 挑戦的萌芽研究
研究期間 : 2014~2015
課題番号 : 26670432
研究分野 : 腎臓内分泌代謝
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