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KAKEN_24591211seika.pdf
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:165.2 KB
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:Apr 11, 2016 |
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ES/iPS細胞の腎尿細管への分化誘導系を用いた腎疾患進展機構の解明
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ES/iPS サイボウ ノ ジン ニョウサイカン エノ ブンカ ユウドウケイ オ モチイタ ジンシッカン シンテン キコウ ノ カイメイ
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ES/iPS saibo no jin nyosaikan eno bunka yudokei o mochiita jinshikkan shinten kiko no kaimei
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An investigation of mechanism of progression of kidney disease using renal tubular cells differentiated from ES/iPS cells
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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・教授
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Research team head
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科研費研究者番号 : 80286484
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2015
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科学研究費補助金研究成果報告書
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2014
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マウスES細胞に胚様体を形成させ, activinとIGFを添加して培養した後, KSP抗体を用いて, KSP陽性細胞を分離することで腎尿細管細胞へ分化誘導を行った。KSP陽性細胞はMatrigel上で管腔を形成し, Wnt4刺激下で尿細管の各セグメントの性質を獲得した。ヒトES細胞はマウスより分化しにくいが, GSK-3β阻害剤によってKSP陽性細胞を増やすことができた。
EMTと上皮化の実験モデルのmicroarray解析から, マウス尿細管細胞においてmiR-34cがTGF-βによるEMTを抑制することがわかった。miR-34cを片側尿管結紮モデルマウスに投与することで腎臓の線維化が抑制された。
We developed a method of inducing renal tubular cells from mouse embryonic stem cells via the cell purification of kidney specific protein (KSP)-positive cells using an anti-KSP antibody. KSP-positive cells had the capacity to form tubular structures when grown in a 3D culture in Matrigel. Moreover, KSP-positive cells acquired the characteristics of each segment of renal tubular cells through tubular formation when stimulated with Wnt4. Human ES cells were not willing to differentiate into tubular cells compared to mouse ES cells. GSK-3ß ; -inhibitor increased KSP-positive cells.
We examined miRNA expression in experimental models of EMT and renal epithelialization using microarray, and found that miR-34c attenuates EMT induced by TGF-β in a mouse tubular cell line. To confirm the effects of miR-34c in vivo, we administered the precursor of miR-34c to mice with unilateral ureteral obstruction, and miR-34c decreased kidney fibrosis.
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研究種目 : 基盤研究(C)
研究期間 : 2012～2014
課題番号 : 24591211
研究分野 : 腎臓内科学
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