我々は, TF-inducible hES細胞株から得られたデータのin silico解析から, 腎前駆細胞及び腎臓Organoidへの分化誘導を促す転写因子群を明らかにした。4転写因子をヒトES細胞に導入後, 2日で92%と高効率に腎前駆細胞を誘導することに成功した。さらにこの腎前駆細胞に別の4因子を添加し3次元培養を行う事で, 14日間で足細胞, 近位及び遠位尿細管からなる腎Organoidを分化誘導することに成功した。RNA-seq法での解析では, 腎Organoidはヒト腎臓遺伝子発現プロファイルと類似した遺伝子発現パターンを示し, ヒト腎臓と近い性質を持った腎臓組織が誘導できていることが示された。
We utilized the transcription factor (TF)-inducible human ES lines, analyzed correlation of gene expression response to the induction of human TFs with tissue-specific gene expression in silico, and identified candidate TFs for differentiating towards renal lineages. Based on in-silico analysis, we have identified a set of four TFs that induce nephron progenitor cells (NPCs) and another set of four TFs that help differentiate toward pretubular aggregate-like cells. Two days after the transfection of the first set of four TFs together into hESCs, NPCs were induced efficiently (~92%). Subsequent administration of the second set of four TFs transfection induced differentiation into pretubular aggregate-like cells. On day 14, epithelial characteristic changes and multi-segmented kidney structures containing podocyte, proximal tubules, and distal tubules were observed in 3D cultures. RNA-Seq analysis also showed the global expression profile closely resembled human kidney profile.

研究種目 : 研究活動スタート支援
研究期間 : 2016～2017
課題番号 : 16H07177
研究分野 : 腎臓再生