Temperature-modulated surfaces for recovery of cultured cells

翻译后摘要：聚（N-异丙基丙烯酰胺）（PIPAAm）显示可逆的亲水/疏水性能变化时，温度的变化。我们已经利用这种聚合物的亲水/疏水表面性质改变的新型温敏恢复系统培养细胞。通过简单的温度降低，可以容易地从PIPAAm接枝的培养皿的表面回收培养的细胞。本文对低温处理培养细胞的脱附机理进行了讨论。通过电子束照射将PIPAAm分子引入聚苯乙烯培养皿。采用牛主动脉内皮细胞和大鼠肝细胞，在37 ℃、5%CO_2的潮湿环境中培养。从细胞表面亲水/疏水性变化和细胞代谢变化两个方面研究了温度对培养细胞从PIPAAm移植的培养皿中回收的影响。在一定温度（4-25 ℃）下孵育30分钟后，在25 ℃下孵育5分钟，可获得更高的培养细胞回收率。用叠氮化钠处理培养细胞，降低了温度处理的细胞恢复，这意味着细胞代谢参与了培养细胞的脱离过程。通过较低温度处理获得的细胞在组织培养皿上增殖良好，并且与通过常规胰蛋白酶处理获得的细胞相比，通过从肝细胞分泌白蛋白来判断，保留了高功能性。细胞的汇合培养物也可以通过温度处理恢复。这些结果有力地表明了利用PIPAAm接枝表面的温度响应性亲水/疏水改变的新型细胞回收系统的可行性。

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.