iPS細胞を用いた不整脈研究の条件検討には大量の心筋細胞が必要であるため, スピナーフラスコによる回転浮遊培養系を用いた心筋細胞の大量培養法の開発と効率的な心筋細胞純化精製法の開発を行った。人工ヌクレアーゼを用いて心筋細胞特異的転写因子(NKX2-5)と刺激伝導系遺伝子のプロモーター下に蛍光蛋白をノックインした組換えヒトiPS細胞を作製し, 心筋細胞刺激伝導細胞への分化および蛍光タンパクの発現を確認した。大型動物への不整脈治療のための心筋細胞移植の基礎実験を行い, ブタにテレメトリー送信機を植え込み不整脈の評価系を確立した。
In order to advance the cell therapy for cardiac arrhythmia, massive amount of iPS Cells-derived cardiomyocytes are necessary. Therefor the massive suspension culture system with a spinner flask and efficient purification strategy of cardiomyocytes were invented. Artificial nucleases were used to insert the fluorescent proteins into the specific locus of cardiac-specific transcription factor (NKX2-5) and conduction system specific gene. These cell lines were specifically differentiated and analyzed. Purified cardiomyocytes were transplanted into pigs and the transplantation strategy was evaluated. Cardiac telemetry transmitters were also implanted into pigs and the evaluation system of arrhythmia was established.

研究種目 : 基盤研究(C)(一般)
研究期間 : 2013～2015
課題番号 : 25505004
研究分野 : 循環器内科