Development of thermoresponsive culture dishes modified with bioactive molecules by affinity binding

we developed a novel method to immobilize biomolecules such as growth factors and enzymes onto thermoresponsive poly(IPAAm-co-CIPAAm)-grafted tissue culture polystyrene dish (TCPS) via affinity binding between avidin and biotin. Effects of RGDS peptides with different spacer length on the behavior of cell attachment / detachment were examined. Human umbilical vein endothelial cells (HUVECs) adhered well on G₁₂RGDS-immobilized thermoresponsive TCPS at 37 °C. Moreover, adhered HUVECs were detached after lowering temperature to 20 °C. However, weak adhesion of HUVECs was observed on thermoresponsive TCPS modified with shorter and longer peptides than G₁₂RGDS. Consequently, thermoresponsive TCPS modified with G₁₂RGDS peptides allows cell culture under the serum-free medium and noninvasive cell recovery by lowering temperature.