Department of Mechanical and Aerospace Engineering, University of Florida, P.O. Box 116250, Gainesville, Florida 32611, USA.
Anal Chem. 2012 May 1;84(9):4199-206. doi: 10.1021/ac3005633. Epub 2012 Apr 17.
Circulating tumor cells (CTC) in the peripheral blood could provide important information for diagnosis of cancer metastasis and monitoring treatment progress. However, CTC are extremely rare in the bloodstream, making their detection and characterization technically challenging. We report here the development of an aptamer-mediated, micropillar-based microfluidic device that is able to efficiently isolate tumor cells from unprocessed whole blood. High-affinity aptamers were used as an alternative to antibodies for cancer cell isolation. The microscope-slide-sized device consists of >59,000 micropillars, which enhanced the probability of the interactions between aptamers and target cancer cells. The device geometry and the flow rate were investigated and optimized by studying their effects on the isolation of target leukemia cells from a cell mixture. The device yielded a capture efficiency of ~95% with purity of ~81% at the optimum flow rate of 600 nL/s. Further, we exploited the device for isolating colorectal tumor cells from unprocessed whole blood; as few as 10 tumor cells were captured from 1 mL of whole blood. We also addressed the question of low throughput of a typical microfluidic device by processing 1 mL of blood within 28 min. In addition, we found that ~93% of the captured cells were viable, making them suitable for subsequent molecular and cellular studies.
循环肿瘤细胞(CTC)在外周血液中可以为癌症转移的诊断和治疗进展监测提供重要信息。然而,CTC 在血液中极其罕见,这使得它们的检测和鉴定具有技术挑战性。我们在这里报告了一种基于适体介导的微柱阵列的微流控装置的开发,该装置能够有效地从未处理的全血中分离肿瘤细胞。高亲和力适体被用作癌症细胞分离的抗体替代品。该显微镜载玻片大小的装置由>59,000 个微柱组成,这增加了适体与靶癌细胞之间相互作用的可能性。通过研究其对从细胞混合物中分离靶白血病细胞的影响,研究了装置的几何形状和流速,并对其进行了优化。在最佳流速为 600 nL/s 时,该装置的捕获效率约为 95%,纯度约为 81%。此外,我们利用该装置从未处理的全血中分离结直肠肿瘤细胞;从 1 毫升全血中可捕获多达 10 个肿瘤细胞。我们还通过在 28 分钟内处理 1 毫升血液来解决典型微流控装置通量低的问题。此外,我们发现约 93%的捕获细胞仍然存活,这使得它们适合随后的分子和细胞研究。