Size-selective collection of circulating tumor cells using Vortex technology
Size-selective collection of circulating tumor cells using Vortex technology
复制标题
DOI：
10.1039/c3lc50689d
复制
发表时间：
2014-01-01
期刊：
影响因子：
6.1
通讯作者：
Di Carlo, Dino
中科院分区：
文献类型：
--
作者：
Sollier, Elodie;Go, Derek E.;Di Carlo, Dino
对于癌症预后和治疗管理，迫切需要一种基于血液的低成本替代放射密集型CT和PET成像的方法。来自相对非侵入性抽血的循环肿瘤细胞（CTC）的“液体活检”是特别理想的，因为它们可以定期重复以提供关于癌症的最新分子信息，这也将为个性化治疗开辟关键机会。除了单纯的诊断应用外，CTC也是药物开发和癌症研究的关注对象。在本文中，我们采用了先前介绍的技术，结合使用微尺度涡旋和惯性聚焦，专门用于从血液样品中高纯度提取CTC。首先，我们系统地改变参数，包括通道尺寸和流速，以达到最大捕获效率和纯度的最佳装置。其次，我们验证了用于捕获血液中癌细胞系的最终装置，考虑了几个因素，包括血液稀释的影响、红细胞裂解和细胞变形性，同时证明了细胞活力和对EpCAM表达的独立性。最后，作为概念验证，从患有乳腺癌（N = 4，25-51个CTC/7.5mL）和肺癌（N = 8，23-317个CTC/7.5mL）的患者的血液中成功提取并计数CTC。重要的是，样品是高纯度的，具有有限的白细胞污染（纯度57-94%）。与现有技术相比，这种Vortex方法具有显著的优势，特别是在处理时间（7.5 mL全血20分钟）、样品浓度（收集低至300 μ L的小体积细胞）、适用于各种癌症类型、细胞完整性和纯度方面。我们预计，它的简单性将有助于临床医生和生物学家的广泛采用，他们不仅希望枚举CTC，而且还希望发现新的CTC生物学，如独特的基因突变，囊泡分泌和转移过程中的作用。
A blood-based, low cost alternative to radiation intensive CT and PET imaging is critically needed for cancer prognosis and management of its treatment. "Liquid biopsies" of circulating tumor cells (CTCs) from a relatively non-invasive blood draw are particularly ideal, as they can be repeated regularly to provide up to date molecular information about the cancer, which would also open up key opportunities for personalized therapies. Beyond solely diagnostic applications, CTCs are also a subject of interest for drug development and cancer research. In this paper, we adapt a technology previously introduced, combining the use of micro-scale vortices and inertial focusing, specifically for the high-purity extraction of CTCs from blood samples. First, we systematically varied parameters including channel dimensions and flow rates to arrive at an optimal device for maximum trapping efficiency and purity. Second, we validated the final device for capture of cancer cell lines in blood, considering several factors, including the effect of blood dilution, red blood cell lysis and cell deformability, while demonstrating cell viability and independence on EpCAM expression. Finally, as a proof-of-concept, CTCs were successfully extracted and enumerated from the blood of patients with breast (N = 4, 25-51 CTCs per 7.5 mL) and lung cancer (N = 8, 23-317 CTCs per 7.5 mL). Importantly, samples were highly pure with limited leukocyte contamination (purity 57-94%). This Vortex approach offers significant advantages over existing technologies, especially in terms of processing time (20 min for 7.5 mL of whole blood), sample concentration (collecting cells in a small volume down to 300 mu L), applicability to various cancer types, cell integrity and purity. We anticipate that its simplicity will aid widespread adoption by clinicians and biologists who desire to not only enumerate CTCs, but also uncover new CTC biology, such as unique gene mutations, vesicle secretion and roles in metastatic processes.