Sakuma Morito, Kumashiro Yoshikazu, Nakayama Masamichi, Tanaka Nobuyuki, Haraguchi Yuji, Umemura Kazuo, Shimizu Tatsuya, Yamato Masayuki, Okano Teruo
Department of Physics, Tokyo University of Science.
Institute of Advanced Biomedical Engineering and Science, Tokyo Women's Medical University;
J Vis Exp. 2016 Mar 1(109):e53465. doi: 10.3791/53465.
Thermoresponsive poly(N-isopropylacrylamide) (PIPAAm)-immobilized surfaces for controlling cell adhesion and detachment were fabricated by the Langmuir-Schaefer method. Amphiphilic block copolymers composed of polystyrene and PIPAAm (St-IPAAms) were synthesized by reversible addition-fragmentation chain transfer (RAFT) radical polymerization. A chloroform solution of St-IPAAm molecules was gently dropped into a Langmuir-trough apparatus, and both barriers of the apparatus were moved horizontally to compress the film to regulate its density. Then, the St-IPAAm Langmuir film was horizontally transferred onto a hydrophobically modified glass substrate by a surface-fixed device. Atomic force microscopy images clearly revealed nanoscale sea-island structures on the surface. The strength, rate, and quality of cell adhesion and detachment on the prepared surface were modulated by changes in temperature across the lower critical solution temperature range of PIPAAm molecules. In addition, a two-dimensional cell structure (cell sheet) was successfully recovered on the optimized surfaces. These unique PIPAAm surfaces may be useful for controlling the strength of cell adhesion and detachment.
通过朗缪尔-谢弗法制备了用于控制细胞黏附与脱离的热响应性聚(N-异丙基丙烯酰胺)(PIPAAm)固定化表面。由聚苯乙烯和PIPAAm组成的两亲性嵌段共聚物(St-IPAAms)通过可逆加成-断裂链转移(RAFT)自由基聚合反应合成。将St-IPAAm分子的氯仿溶液缓慢滴入朗缪尔槽装置中,然后水平移动装置的两个挡板以压缩薄膜,调节其密度。接着,通过表面固定装置将St-IPAAm朗缪尔膜水平转移到疏水改性的玻璃基板上。原子力显微镜图像清晰地显示出表面的纳米级海岛结构。通过在PIPAAm分子的低临界溶液温度范围内改变温度,可以调节细胞在制备表面上黏附与脱离的强度、速率和质量。此外,在优化后的表面上成功回收了二维细胞结构(细胞片)。这些独特的PIPAAm表面可能有助于控制细胞黏附与脱离的强度。