Preparation of cell culture dishes immobilized nanobrush for regenerative medicine and preservation and culture of stem cells

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The triblock copolymer Pluronic, composed of polyethylene oxide(PEO, hydrophilic segment)-polypropylene oxide(PPO, hydrophobic segment)-PEO(hydrophilic segment) triblocks, exhibits amphiphilic properties, and is temperature responsive polymer having low critical solution temperature(LCST). The surface having hydrophilic and hydrophilic domains is known to show biocompatibility than the surface having only hydrohilic domain. However, the biocompatible materials having amphiphilic domains in perpendicular to the surface have not yet investigated. In this project, amphiphilic triblock copolymer(pluronic) having several segment length(molecular weight) and amphiphilic content was designed and grafted on cell culture dishes by surface reaction, and thus, the cell culture dishes prepared from amphiphilic triblock copolymer was developed(cell culture dishes having nanosegments). One of the goal of this project is to investigate the optimal molecular design of cell culture dishes having nanose … More gments, which was prepared from amphiphilic triblock copolymers.The second goal of this project is that detachment of the cells cultured on cell culture dishes grafted nanobrush, which is composed of amphiphilic triblock copolymer having LCST, was investigated without using trypsin in cell detachment and that the optimal design of cell culture dishes having nanobrush was investigated. Preservation of hematopoietic stem cells were efficiently possible on the dishes immobilized triblock copolymer having 70% hydrophilic domain and 10, 000 dalton of molecular weight. The dishes having high density of nanobrush were not suitable to culture and preserve the hematopoietic stem cells.Several extracellular matrices(ECMs, gelatin, collagen, laminin, vitronectin, fibronectin, and matrigel) were immobilized on polystyrene dishes by covalent bonding and coating method. The surface marker of mesenchymal stem cells(MSCs) cultured on different dishes immobilized ECMs show comparable expression. MSCs cultured on gelatin and vitronectin-coating dishes tended to differentiate into osteoblasts in differentiation medium. However, MSC cultured on ECM-grafted dishes did not promote to differentiate into osteoblasts. MSCs cultured on ECM-grafted dishes were found to promote differentiation into neural cells. Especially, MSCs cultured on matrigel-and laminin-grafted dishes showed higher differentiation into neural cells. MSCs cultured on ECM and pluronic grafted dishes can be detached from the dishes by decreasing temperature of dishes at 4℃ Less