Pugia Michael, Magbanua Mark Jesus M, Park John W
Siemens Healthcare Diagnostics, 3400 Middlebury Street, Elkhart, IN, 46515, USA.
Division of Hematology & Oncology, University of California San Francisco, San Francisco, CA, 94115, USA.
Adv Exp Med Biol. 2017;994:119-131. doi: 10.1007/978-3-319-55947-6_6.
The current standard methods for isolating circulating tumor cells (CTCs) from blood involve EPCAM-based immunomagnetic approaches. A major disadvantage of these strategies is that CTCs with low EPCAM expression will be missed. Isolation by size using filter membranes circumvents the reliance on this cell surface marker, and can facilitate the capture not only of EPCAM-negative CTCs but other rare cells as well. These cells that are trapped on the filter membrane can be characterized by immunocytochemistry (ICC) , enumerated and profiled to elucidate their clinical significance. In this chapter, we discuss advances in filtration systems to capture rare cells as well as downstream ICC methods to detect and identify these cells. We highlight our recent clinical study demonstrating the feasibility of using a novel method consisting of automated microfluidic filtration and sequential ICC for detection and enumeration of CTCs, as well as circulating mesenchymal cells (CMCs), circulating endothelial cells (CECs), and putative circulating stem cells (CSCs). We hypothesize that simultaneous analysis of circulating rare cells in blood of cancer patients may lead to a better understanding of disease progression and development of resistance to therapy.
目前从血液中分离循环肿瘤细胞（CTC）的标准方法涉及基于上皮细胞粘附分子（EPCAM）的免疫磁珠法。这些策略的一个主要缺点是低EPCAM表达的CTC会被遗漏。使用滤膜按大小进行分离避免了对这种细胞表面标志物的依赖，不仅可以捕获EPCAM阴性的CTC，还能捕获其他稀有细胞。捕获在滤膜上的这些细胞可以通过免疫细胞化学（ICC）进行表征、计数和分析，以阐明其临床意义。在本章中，我们讨论了用于捕获稀有细胞的过滤系统的进展以及用于检测和识别这些细胞的下游ICC方法。我们重点介绍了我们最近的一项临床研究，该研究证明了使用一种由自动微流控过滤和顺序ICC组成的新方法来检测和计数CTC以及循环间充质细胞（CMC）、循环内皮细胞（CEC）和假定的循环干细胞（CSC）的可行性。我们假设对癌症患者血液中的循环稀有细胞进行同步分析可能有助于更好地理解疾病进展和耐药性的产生。
