High-purity and label-free isolation of circulating tumor cells (CTCs) in a microfluidic platform by using optically-induced-dielectrophoretic (ODEP) force
High-purity and label-free isolation of circulating tumor cells (CTCs) in a microfluidic platform by using optically-induced-dielectrophoretic (ODEP) force
复制标题
DOI：
10.1039/c3lc41256c
复制
发表时间：
2013-01-01
期刊：
影响因子：
6.1
通讯作者：
Lee, Gwo-Bin
中科院分区：
文献类型：
--
作者：
Huang, Song-Bin;Wu, Min-Hsien;Lee, Gwo-Bin
基于阴性选择的循环肿瘤细胞(CTC)分离被认为是有价值的，以获得更多的天然CTC，特别是所有可能的CTC，而不存在与CTC表面抗原特性相关的偏见。然而，在这样的细胞分离策略下，CTC的纯度通常会受到影响。为了解决这个问题，这项研究报告了基于光诱导-介电泳法(ODEP)的细胞操纵和微流控平台中的层流模式的集成，用于在传统的基于阴性选择的CTC分离之后分离未经处理的高纯度CTC。在设计中，根据癌细胞和白细胞的大小差异和电学性质，在两个平行的层流中连续同时施加六段移动的光条屏，以同时分离癌细胞和白细胞。分离的细胞群通过这两个流进一步划分、传递和收集。通过这种方法，可以以连续、有效和高效的方式分离癌细胞。在本研究中，首次对ODEP操作前列腺癌(PC-3)和口腔癌(OEC-M1)细胞以及有少量细胞聚集现象的白细胞的操作条件进行了表征。此外，还对该方法分离癌细胞的性能进行了实验研究。结果表明，该分离方案能够从白细胞背景中分离PC-3细胞或OEC-M1细胞，回收率高(PC-3细胞：76-83%，OEC-M1细胞：61-68%)，纯度高(PC-3细胞：74-82%，OEC-M1细胞：-66%)(设定流速：0.1亩L min(-1)，进样量：1ml)。后者超出了目前在常规四氯化碳隔离中可能实现的范围。此外，分离的癌细胞对PC-3和OEC-M1细胞的存活率分别高达94+/-2%和95+/-3%。此外，分离的癌细胞也被证明保持了它们的增殖能力。作为一个整体，本研究提出了一种基于ODEP的微流控平台，能够以连续的、无标记的、细胞友好的、特别是高纯度的方式分离CTCs。所有这些特征都被发现对于利用收获的CTCs进行后续的基于细胞的或生化分析特别有意义。
Negative selection-based circulating tumor cell (CTC) isolation is believed valuable to harvest more native, and in particular all possible CTCs without biases relevant to the properties of surface antigens on the CTCs. Under such a cell isolation strategy, however, the CTC purity is normally compromised. To address this issue, this study reports the integration of optically-induced-dielectrophoretic (ODEP) force-based cell manipulation, and a laminar flow regime in a microfluidic platform for the isolation of untreated, and highly pure CTCs after conventional negative selection-based CTC isolation. In the design, six sections of moving light-bar screens were continuously and simultaneously exerted in two parallel laminar flows to concurrently separate the cancer cells from the leukocytes based on their size difference and electric properties. The separated cell populations were further partitioned, delivered, and collected through the two flows. With this approach, the cancer cells can be isolated in a continuous, effective, and efficient manner. In this study, the operating conditions of ODEP for the manipulation of prostate cancer (PC-3) and human oral cancer (OEC-M1) cells, and leukocytes with minor cell aggregation phenomenon were first characterized. Moreover, performances of the proposed method for the isolation of cancer cells were experimentally investigated. The results showed that the presented CTC isolation scheme was able to isolate PC-3 cells or OEC-M1 cells from a leukocyte background with high recovery rate (PC-3 cells: 76-83%, OEC-M1 cells: 61-68%), and high purity (PC-3 cells: 74-82%, OEC-M1 cells: 64-66%) (set flow rate: 0.1 mu l min(-1) and sample volume: 1 ml). The latter is beyond what is currently possible in the conventional CTC isolations. Moreover, the viability of isolated cancer cells was evaluated to be as high as 94 +/- 2%, and 95 +/- 3% for the PC-3, and OEC-M1 cells, respectively. Furthermore, the isolated cancer cells were also shown to preserve their proliferative capability. As a whole, this study has presented an ODEP-based microfluidic platform that is capable of isolating CTCs in a continuous, label-free, cell-friendly, and particularly highly pure manner. All these traits are found particularly meaningful for exploiting the harvested CTCs for the subsequent cell-based, or biochemical assays.