The Bradley Department of Electrical and Computer Engineering , Virginia Tech , Blacksburg , Virginia 24061 , United States.
Department of Cancer Biology , Wake Forest University School of Medicine , Winston-Salem , North Carolina 27157 , United States.
Anal Chem. 2018 Jun 19;90(12):7526-7534. doi: 10.1021/acs.analchem.8b01134. Epub 2018 Jun 1.
Circulating tumor cells (CTCs) are broadly accepted as an indicator for early cancer diagnosis and disease severity. However, there is currently no reliable method available to capture and enumerate all CTCs as most systems require either an initial CTC isolation or antibody-based capture for CTC enumeration. Many size-based CTC detection and isolation microfluidic platforms have been presented in the past few years. Here we describe a new size-based, multiple-row cancer cell entrapment device that captured LNCaP-C4-2 prostate cancer cells with >95% efficiency when in spiked mouse whole blood at ∼50 cells/mL. The capture ratio and capture limit on each row was optimized and it was determined that trapping chambers with five or six rows of micro constriction channels were needed to attain a capture ratio >95%. The device was operated under a constant pressure mode at the inlet for blood samples which created a uniform pressure differential across all the microchannels in this array. When the cancer cells deformed in the constriction channel, the blood flow temporarily slowed down. Once inside the trapping chamber, the cancer cells recovered their original shape after the deformation created by their passage through the constriction channel. The CTCs reached the cavity region of the trapping chamber, such that the blood flow in the constriction channel resumed. On the basis of this principle, the CTCs will be captured by this high-throughput entrapment chip (CTC-HTECH), thus confirming the potential for our CTC-HTECH to be used for early stage CTC enrichment and entrapment for clinical diagnosis using liquid biopsies.
循环肿瘤细胞 (CTC) 被广泛认为是癌症早期诊断和疾病严重程度的指标。然而,目前还没有可靠的方法来捕获和计数所有的 CTC,因为大多数系统要么需要初始 CTC 分离,要么需要抗体捕获来进行 CTC 计数。过去几年已经提出了许多基于大小的 CTC 检测和分离微流控平台。在这里,我们描述了一种新的基于大小的、多排癌细胞捕获装置,当在约 50 个细胞/mL 的掺入小鼠全血中时,该装置能够以 >95%的效率捕获 LNCaP-C4-2 前列腺癌细胞。优化了每排的捕获比和捕获极限,确定需要 5 排或 6 排微收缩通道的捕获室才能达到 >95%的捕获比。该装置在入口处以恒定压力模式操作血液样本,在该阵列中的所有微通道中产生均匀的压力差。当癌细胞在收缩通道中变形时,血流暂时减慢。一旦进入捕获室,癌细胞在通过收缩通道产生的变形后恢复其原始形状。CTC 到达捕获室的腔区域,使得收缩通道中的血流恢复。基于这一原理,CTC 将被这种高通量捕获芯片(CTC-HTECH)捕获,从而证实了我们的 CTC-HTECH 有潜力用于临床诊断中使用液体活检进行早期 CTC 富集和捕获。