| dc.description.abstract | Human pluripotent stem cells (hPSCs) can be divided into human embryonic stem cells (hESCs) and human induced pluripotent stem cells (hiPSCs), which can differentiate into the cells derived from three germ layers, such as endoderm, ectoderm, and mesoderm. Typically, conventional tissue culture polystyrene plates cannot support the proliferation of hPSCs, unlike most cancer cells or tissue cells that can proliferate. Alternatively, hPSCs are commonly cultured on dishes coated with Matrigel, which involves xeno-containing substances. Hence, it is advisable to employ biomaterials grafted with synthetic peptides to facilitate hPSC adhesion, maintain hPSC pluripotency, and differentiate hPSCs for the clinical application of hPSCs. The specific peptide-grafted PVA-IA (poly (vinyl alcohol-co-vinyl acetate-co-itaconic acid)) hydrogels with optimal elasticity (25.3 kPa) have been developed to support the proliferation of hPSCs, where the hydrogels grafted with specific oligopeptides derived from laminin-β4 (LMN) and vitronectin were the most preferable biomaterials for hPSC proliferation and could be used as cell culture biomaterials of hPSCs for more than 10 passages. However, the concentration of oligopeptide solution needed for grafting on the hydrogels should be much higher (typically 1000 μg/mL) than the concentration of ECM proteins used for the preparation of ECM protein-coated surface (typically 5-10 μg/mL). To improve this point, a new design of cell culture biomaterials using a dendrimer-based peptide-grafted surface was developed in this study. Polyamidoamine (PAMAM) dendrimer having generation 3, which was composed of many branched subunits of amide and amine groups, was used to be immobilized on the hydrogel surface, where it is expected to have high biocompatibility for hPSC culture and differentiation. The cell proliferation of hiPSCs on PAMAM dendrimer surface grafted with CLB1GK with different peptide concentration were evaluated from expansion fold. It is evaluated which peptide concentration was more suitable for hiPSC proliferation in this study. The hPSCs could be cultured on PAMAM dendrimer surface grafted with the peptide, which were prepared with low concentration of peptide (50 μg/mL) and where the concentration of peptide is the same order of the concentration of coating ECM solution (5-10 μg/mL) on ECM protein-coated surface. The hiPSCs showed their pluripotency and differentiation ability into three germ layer cells after long-term (10 passage) culture on the peptide-grafted PAMAM dendrimer hydrogel surface, which were prepared with a low peptide concentration (50 μg/ml). Currently, only this study supports hPSC proliferation on the peptide-grafted surface, which was prepared with a low concentration of peptide solution (50 μg/ml), whereas most typical peptide-grafted biomaterials for hPSC culture was prepared with a high concentration of peptide solution, such as >1000 μg/ml. 3D peptide location (immobilization) on the surface using PAMAM dendrimer enables to maintain the pluripotency of hPSCs during their cultivation for
a long time. | en_US |