Differentiation of Induced Pluripotent Stem Cells in TATVHL Peptide and PLGA Nanoparticles-grafted Three-dimensionally Ordered Chitosan-PAAM Scaffolds

• Main Authors: Chun-Wei, Chen, 陳俊瑋
• Other Authors: 郭勇志
• Format: Others
• Language: zh-TW
• Published: 2013
• Online Access: http://ndltd.ncl.edu.tw/handle/52796943811236929477

碩士 === 國立中正大學 === 化學工程研究所 === 101 === In this research, we manufactured three-dimensionally chitosan-polyacrylamide scaffolds by inverted colloidal crystals method. The pore surfaces were grafted with poly(lactide-co-glycolide) nanoparticles for surface roughness by EDC and NHS, and grafted with TATVHL peptide by BS-3 crosslinking agent. When mass ratio of chitosan/polyacrylamide = 1:1 is the best for scaffold composition. Adhesion efficiency of iPS cells in TATVHL peptide-grafted (15 μg/ml) scaffold increased 9.8% compared to unmodified scaffold. Adhesion efficiency of iPS cells in PLGA nanoparticles-grafted (1.5 mg/ml)scaffold increase 3.8% compared to unmodified scaffold. TATVHL peptide-grafted scaffold containing seeded iPS cells cultured for 7 days, the SEM images of scaffold indicated TATVHL peptide-grafted scaffold guide iPS cells to differentiate toward neurons. PLGA nanoparticle and TATVHL peptide-grafted scaffold guided iPS cells to differentiate toward neurons significantly, the neurofilament was about 15 μm. Immunohistochemical fluorescence staining indicated TATVHL peptide could inhibit iPS cells differentiate to astrocytes, and TATVHL peptide could guide iPS cells to differentiate toward neurons. The flow cytometry analysis indicated that the III tubulin-positive cells were proportional to the amount of TATVHL peptide. The percentage of III tubulin increased 30-60% of TATVHL peptide-grafted scaffolds compared to unmodified scaffold. PLGA nanoparticle and TATVHL peptide-grafted scaffold guided iPS cells to differentiate toward neurons significantly compared to TATVHL peptide-grafted scaffold.