Department of Power Mechanical Engineering, National Tsing Hua University, Hsinchu, Taiwan.
National Institute of Cancer Research, National Health Research Institutes, Tainan, Taiwan.
Sci Rep. 2017 Jun 26;7(1):4248. doi: 10.1038/s41598-017-04056-2.
Cholangiocarcinoma (CCA), a biliary tract malignancy, accounts for 20% of all liver cancers. There are several existing methods for diagnosis of CCA, though they are generally expensive, laborious, and suffer from low detection rates. Herein we first developed a means of partially purifying human bile for consequent injection into a microfluidic chip. Then, the novel microfluidic system, which featured 1) a cell capture module, 2) an immunofluorescence (IF) staining module featuring two CCA-specific biomarkers, and 3) an optical detection module for visualization of antibody probes bound to these CCA marker proteins, was used to detect bile duct cancer cells within partially purified bile samples. As a proof of concept, CCA cells were successfully captured and identified from CCA cell cultures, blood samples inoculated with CCA cells, and clinical bile specimens. In 7.5 ml of bile, this system could detect >2, 0, and 1 positive cells in advanced stage patients, healthy patients, and chemotherapy-treated patients, respectively. In conclusion, our microfluidic system could be a promising tool for detection of cancer cells in bile, even at the earliest stages of CCA when cancer cells are at low densities relative to the total population of epithelial cells.
胆管癌（CCA）是一种胆道恶性肿瘤，占所有肝癌的 20%。目前有几种现有的 CCA 诊断方法，但它们通常昂贵、繁琐，并且检测率较低。在此，我们首次开发了一种部分纯化人胆汁的方法，以便随后注入微流控芯片。然后，使用新型微流控系统，其具有 1）细胞捕获模块、2）具有两种 CCA 特异性生物标志物的免疫荧光（IF）染色模块，以及 3）用于可视化与这些 CCA 标记蛋白结合的抗体探针的光学检测模块，来检测部分纯化的胆汁样本中的胆管癌细胞。作为概念验证，我们成功地从 CCA 细胞培养物、接种 CCA 细胞的血液样本和临床胆汁标本中捕获和鉴定了 CCA 细胞。在 7.5ml 的胆汁中，该系统可以分别检测到晚期患者、健康患者和化疗治疗患者中的>2、0 和 1 个阳性细胞。总之，我们的微流控系统可能是一种有前途的工具，可用于检测胆汁中的癌细胞，即使在 CCA 的最早阶段，当癌细胞相对于上皮细胞的总群体密度较低时也是如此。
