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KAKEN_25461231seika.pdf
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Title |
Title |
ヒトES・iPS細胞から腎尿細管細胞への分化誘導技術の確立および医薬応用
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ヒト ES・iPS サイボウ カラ ジン ニョウサイカン サイボウ エノ ブンカ ユウドウ ギジュツ ノ カクリツ オヨビ イヤク オウヨウ
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Hito ES・iPS saibo kara jin nyosaikan saibo eno bunka yudo gijutsu no kakuritsu oyobi iyaku oyo
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Generation of kidney tubular cells from human pluripotent stem cells
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本間, 康一郎
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ホンマ, コウイチロウ
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Honma, Koichiro
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慶應義塾大学・医学部・講師
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Research team head
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科研費研究者番号 : 10383762
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山口, 慎太郎
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ヤマグチ, シンタロウ
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Yamaguchi, Shintaro
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慶應義塾大学・医学部・特任助教
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Research team member
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科研費研究者番号 : 50464855
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2017
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科学研究費補助金研究成果報告書
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2016
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Abstract |
3日間GSK3β阻害剤を用いて, hPSCを中胚葉細胞に分化させ, 次に, これらの細胞を腎上皮増殖培地で培養し, KSP+細胞を誘導した。抗KSP抗体によるフローサイトメトリーによりKSP+細胞を純化すると, それらの細胞は腎尿細管細胞のあらゆるセグメントの特徴を示し, またKSP+細胞を3Dマトリゲルで培養するとin vitroで尿細管オルガノイドを形成した。KSP+細胞による尿細管オルガノイドの形成は機能的な腎尿細管の獲得を誘導した。また, KSP+細胞は, マウス胚性腎臓細胞とともに培養すると, 3D尿細管構造内で自己組み立てを行うので, ヒトとマウスのキメラ腎臓培養物の作製が可能であった。
We report a simple two-step differentiation protocol to generate kidney tubular organoids from hPSCs with direct purification of KSP (kidney specific protein)-positive cells using anti-KSP antibody. We first differentiated hPSCs into mesoderm cells using a glycogen synthase kinase-3β inhibitor for 3 days, then cultured cells in renal epithelial growth medium to induce KSP+ cells. We purified KSP+ cells using flow cytometry with anti-KSP antibody, which exhibited characteristics of all segments of kidney tubular cells and cultured KSP+ cells in 3D Matrigel, which formed tubular organoids in vitro. The formation of tubular organoids by KSP+ cells induced the acquisition of functional kidney tubules. KSP+ cells also allowed for the generation of chimeric kidney cultures in which human cells self-assembled into 3D tubular structures in combination with mouse embryonic kidney cells.
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研究種目 : 基盤研究(C)(一般)
研究期間 : 2013~2016
課題番号 : 25461231
研究分野 : 腎臓内科学
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