Temperature-modulated surfaces for recovery of cultured cells

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Abstract :Poly (N-isopropylacrylamide) (PIPAAm) shows reversible hydrophilic / hydrophobic property changes upon temperature changes. We have utilized this polymer for hydrophilic / hydrophobic surface property alteration for novel thermo-responsive recovery system for cultured cells. Cultured cells can easily be recovered from the surfaces of PIPAAm-grafted culture dishes by simple temperature decreases. In this report detachment mechanism of cultured cells by lowering temperature treatment is discussed. PIPAAm molecules are introduced to polystirene culture dishes by electron-beam irradiation. Used cells are bovine aortic endothelial cells and rat hepatocytes and cultured in a humidified atmosphere of 5% CO_2 at 37ﾟC.The effect of temperature on recovery of cultured cells from PIPAAm-grafted dish was investigated with respect to both degree of surface hydrophilic / hydrophobic change and cell metabolism change. Greater recovery of cultured cells can be achieved by incubation at 25ﾟC for 5 min after 30 min incubation at a definite temperature (4-25ﾟC). Recovery of cells by temperature treatment was decreased by treating cultured cells with sodium azide, implying that cellular metabolism is involved in the detachment process of cultured cells. Recovered cells by lower temperature treatment well proliferated on tissue culture dishes and retained high functionality as judged by secretion of albumin from hepatocytes compared with cells recovered by conventional trypsin treatment. Confluent culture of cells could also be recovered by temperature treatment. This technique might be useful for construction of artificial tissues such as artificial skin.These results strongly suggest the feasibility of novel cell recovery system using temperature-responsive hydrophilic / hydrophobic alterations of PIPAAm-grafted surfaces.