Project/Area Number |
26461185
|
Research Category |
Grant-in-Aid for Scientific Research (C)
|
Allocation Type | Multi-year Fund |
Section | 一般 |
Research Field |
Respiratory organ internal medicine
|
Research Institution | The University of Tokyo |
Principal Investigator |
SAITO Akira 東京大学, 保健・健康推進本部, 助教 (90591412)
|
Research Collaborator |
NAGASE Takahide
OHSHIMA Mitsuhiro
YAMAGUCHI Yoko
SUZUKI Hiroshi
HORIE Masafumi
NOGUCHI Satoshi
OKUNISHI Katsuhide
ONO Naoaki
MICKE Patrick
KACZKOWSKI Bogumil
|
Project Period (FY) |
2014-04-01 – 2017-03-31
|
Project Status |
Completed (Fiscal Year 2016)
|
Budget Amount *help |
¥4,940,000 (Direct Cost: ¥3,800,000、Indirect Cost: ¥1,140,000)
Fiscal Year 2016: ¥910,000 (Direct Cost: ¥700,000、Indirect Cost: ¥210,000)
Fiscal Year 2015: ¥910,000 (Direct Cost: ¥700,000、Indirect Cost: ¥210,000)
Fiscal Year 2014: ¥3,120,000 (Direct Cost: ¥2,400,000、Indirect Cost: ¥720,000)
|
Keywords | リプログラミング / 遺伝子導入 / 肺上皮細胞 / 肺線維芽細胞 / 小細胞肺癌 / 非小細胞肺癌 / 肺線維症 / トランスクリプトーム / 転写因子 / 肺胞上皮細胞 / トランスクリプト―ム / 転写ネットワーク / 三次元培養 / 動物モデル |
Outline of Final Research Achievements |
We explored optimal settings for gene transduction by electroporation in human iPS cells and dermal fibroblasts, and determined the condition for low cytotoxicity and high transduction efficiency. We constructed expression vectors for TTF-1, YAP/TAZ, and FOXA2, and attempted to induce cellular differentiation via ectopic expression of these factors. We also tested the effects of TGF-beta/BMP-4/FGF-2 ligands/inhibitors, demethylating agent, and HDAC inhibitor. In an attempt to establish suitable conditions for lung epithelial cell differentiation, serum-free media, collagen gels embedded with lung fibroblasts, three-dimensional and air-liquid interface cultures were utilized. Elevated expression of lung epithelial cell markers (SPC, CC10, and FoxJ1) were confirmed by quantitative PCR, suggesting that lung epithelial cell differentiation with a certain level was achieved through combinations of gene transfer and culture methods.
|