Department of Mechanical Engineering, Columbia University, New York, NY 10027, USA.
Lab Chip. 2012 Sep 21;12(18):3504-13. doi: 10.1039/c2lc40411g. Epub 2012 Aug 2.
Isolation of cells from heterogeneous mixtures is critically important in both basic cell biology studies and clinical diagnostics. Cell isolation can be realized based on physical properties such as size, density and electrical properties. Alternatively, affinity binding of target cells by surface-immobilized ligands, such as antibodies, can be used to achieve specific cell isolation. Microfluidics technology has recently been used in conjunction with antibody-based affinity isolation methods to capture, purify and isolate cells with higher yield rates, better efficiencies and lower costs. However, a method that allows easy release and collection of live cells from affinity surfaces for subsequent analysis and detection has yet to be developed. This paper presents a microfluidic device that not only achieves specific affinity capture and enrichment, but also enables non-destructive, temperature-mediated release and retrieval of cells. Specific cell capture is achieved using surface-immobilized aptamers in a microchamber. Release of the captured cells is realized by a moderate temperature change, effected via integrated heaters and a temperature sensor, to reversibly disrupt the cell-aptamer interaction. Experimental results with CCRF-CEM cells have demonstrated that the device is capable of specific capture and temperature-mediated release of cells, that the released cells remain viable and that the aptamer-functionalized surface is regenerable.
从异质混合物中分离细胞在基础细胞生物学研究和临床诊断中都至关重要。细胞分离可以基于物理性质(如大小、密度和电特性)来实现。或者,可以使用表面固定的配体(如抗体)与靶细胞的亲和结合来实现特定的细胞分离。微流控技术最近已与基于抗体的亲和分离方法结合使用,以提高产量、效率和降低成本来捕获、纯化和分离细胞。然而,仍然需要开发一种能够轻松地从亲和表面上释放和收集活细胞以进行后续分析和检测的方法。本文提出了一种微流控设备,不仅能够实现特异性亲和捕获和富集,还能够实现非破坏性、温度介导的细胞释放和回收。使用微腔中的表面固定适体实现特定细胞的捕获。通过集成的加热器和温度传感器实现温和的温度变化来实现捕获细胞的释放,从而可逆地破坏细胞-适体相互作用。用 CCRF-CEM 细胞进行的实验结果表明,该设备能够实现细胞的特异性捕获和温度介导的释放,释放的细胞保持活力,并且适体功能化表面是可再生的。