Construction of cell culture system by combining cell design using transfection of genes and material design using immobilization of biological molecules

对于再生医学，需要在没有血清或动物来源饲养细胞的情况下进行细胞培养。1)设计并制备了共表达促红细胞生成素和表皮生长因子受体的中国仓鼠卵巢细胞。将细胞接种于EGF固定的培养板上。在固定化EGF的存在下，细胞生长速度加快，分泌的促红细胞生成素增加。2)采用梯度微区固定化EGF、肝素和温敏聚合物。高密度固定化EGF才能显著促进高表达EGF受体的CHO细胞的生长。NIH3T3细胞的生长只有在高密度的固定化肝素和碱性成纤维细胞生长因子存在的情况下才能被促进。认为碱性成纤维细胞生长因子是在肝素固定区被激活的。培养在温度响应性聚合物固定化表面上的细胞的分离受到温度控制的调节。此外，剥离还依赖于梯度模式。梯度微图案固定化被认为是研究固定化分子效果的重要工具。

For regenerative medicine, cell culture without serum or feeder cells from animal sources is needed. In the present study, cell culture system was constructed using cell design by gene transfection and material design by immobilization of biosignal molecules.1) Chinese hamster ovary cell coexpressiong erythropoietin and epidermal growth factor (EGF) receptor was designed and prepared. The cell was cultured on the plate immobilized with EGF. The growth rate was highly accelerated in the presence of immobilized EGF and the amount of secreted erythropoietin was increased.2) Gradient-micropattern immobilization of EGF, heparin, and thermo-responsive polymers was carried out. Growth of CHO cells overexpressing EGF receptor was significantly enhanced only on the high density of immobilized EGF. The growth of NIH3T3 cells was enhanced only on the high density of immobilized heparin in the presence of basic fibroblast growth factor (bFGF). It was considered that bFGF was activated on the heparin-immobilized regions. Detachment of cells cultured on the thermo-responsive polymer-immobilized surface was regulated by temperature control. In addition, the detachment was dependent on the gradient pattern. The gradient micropattern immobilization is considered to an important tool for investigation of the effect of immobilized molecules.