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'Smart' biopolymer for a reversible stimuli-responsive platform in cell-based biochips

机译:用于基于细胞的生物芯片中可逆的刺激反应平台的“智能”生物聚合物

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The rapid response of a smart material surface to external stimuli is critical for application to cell-based biochips. The sharp and controllable phase transition of elastin-like polypeptide (ELP) enabled reversible cell adhesion on the surface by changing the temperature or salt concentration in the system. First, ELP micropatterns were prepared on a glass surface modified into aldehyde. The lysine-containing ELP (ELP-K) was genetically synthesized from E. coli for conjugation with the aldehyde on the glass surface. The phase transition of ELP was monitored in PBS and cell culture media using UV-visible spectroscopy, and a significant difference in transition temperature (T-t) was observed between the two solution systems. The micropatterning of ELP on the glass surface was performed by microcontact printing a removable polymeric template on the aldehyde-glass followed by incubation in ELP-K aqueous solution. The ELP micropatterns were imaged with atomic force microscopy and showed a monolayer thickness of similar to 4 nm. Imaging from time-of-flight secondary ion mass spectroscopy confirmed that the ELP molecules were successfully immobilized on the highly resolved micropatterns. Cell attachment and detachment could be reversibly controlled on the ELP surfaces by external stimuli. The hydrophobic phase above T-t resulted in the adhesion of fibroblasts, while the detachment of cells was induced by lowering the incubation temperature below T-t. The smart properties of ELP were reliable and reproducible, demonstrating potential applications in cell-based microdevices.
机译:智能材料表面对外部刺激的快速响应对于应用于基于细胞的生物芯片至关重要。弹性蛋白样多肽(ELP)的尖锐且可控的相变可通过改变系统中的温度或盐浓度来实现表面上可逆的细胞粘附。首先,在改性为醛的玻璃表面上制备ELP微图案。含赖氨酸的ELP(ELP-K)是从大肠杆菌中遗传合成的,用于与玻璃表面的醛结合。使用紫外可见光谱法在PBS和细胞培养基中监测ELP的相变,并且观察到两种溶液系统之间的转变温度(T-t)有显着差异。通过在醛玻璃上微接触印刷可移除的聚合物模板,然后在ELP-K水溶液中孵育,在玻璃表面上进行ELP的微图案化。用原子力显微镜对ELP微型图案进行成像,显示单层厚度接近4 nm。飞行时间二次离子质谱的成像证实,ELP分子已成功固定在高度分辨的微图案上。细胞的附着和分离可以通过外部刺激可逆地控制在ELP表面。 T-t上方的疏水相导致成纤维细胞的粘附,而将温育温度降低到T-t以下则诱导了细胞的脱离。 ELP的智能特性可靠且可重复,证明了在基于细胞的微型设备中的潜在应用。

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