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

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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.